Tetrahydro-Pyrido-Pyrimidine Derivatives

ABSTRACT

The invention relates to substituted tetrahydro-pyrido-pyrimidine derivatives of the formula (I), 
     
       
         
         
             
             
         
       
     
     wherein Y, R 1 , R 2  and m are as defined in the description. Such compounds are suitable for the treatment of a disorder or disease which is mediated by the activity of the PI3K enzymes.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.15/285,614, filed Oct. 5, 2016, the entire contents of which is herebyincorporated herein by reference. U.S. application Ser. No. 15/285,614,is a continuation of U.S. application Ser. No. 14/151,796, filed Jan. 9,2014, now U.S. Pat. No. 9,499,536, the entire contents of which ishereby incorporated herein by reference. U.S. application Ser. No.14/151,796, is a divisional of U.S. patent application Ser. No.13/175,050, filed Jul. 1, 2011, now U.S. Pat. No. 8,653,092, the entirecontents of which is hereby incorporated herein by reference. U.S.application Ser. No. 13/175,050, claims the benefit of U.S. ProvisionalPatent Application No. 61/361,589, filed Jul. 6, 2010, the entirecontents of which is hereby incorporated herein by reference

FIELD OF THE INVENTION

The invention relates to the preparation and use of newtetrahydro-pyrido-pyrimidine derivatives as drug candidates in free formor in pharmaceutically acceptable salt form with valuable druglikeproperties, such as e.g. metabolic stability and suitablepharmacokinetics, form for the modulation, notably the inhibition of theactivity or function of the phosphoinositide 3′ OH kinase family(hereinafter PI3K).

BACKGROUND OF THE INVENTION

The invention relates to the treatment, either alone or in combination,with one or more other pharmacologically active compounds, ofPI3K-related diseases including but not limited to autoimmune disorders,inflammatory diseases, allergic diseases, airway diseases, such asasthma and COPD, transplant rejection, cancers eg of hematopoieticorigin or solid tumors. The invention also relates to the treatment,either alone or in combination, with one or more other pharmacologicallyactive compounds, includes methods of treating conditions, diseases ordisorders in which one or more of the functions of B cells such asantibody production, antigen presentation, cytokine production orlymphoid organogenesis are abnormal or are undesirable includingrheumatoid arthritis, pemphigus vulgaris, idiopathic thrombocytopeniapurpura, systemic lupus erythematosus, multiple sclerosis, myastheniagravis, Sjögren's syndrome, autoimmune hemolytic anemia, ANCA-associatedvasculitides, cryoglobulinemia, thrombotic thrombocytopenic purpura,chronic autoimmune urticaria, allergy (atopic dermatitis, contactdermatitis, allergic rhinitis), goodpasture's syndrome, AMR(antibody-mediated transplant rejection), B cell-mediated hyperacute,acute and chronic transplant rejection and cancers of haematopoieticorigin including but not limited to multiple myeloma; acute myelogenousleukemia; chronic myelogenous leukemia; lymphocytic leukemia; myeloidleukemia; non-Hodgkin lymphoma; lymphomas; polycythemia vera; essentialthrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenstroem disease.

SUMMARY OF THE INVENTION

In a first aspect, the invention relates to tetrahydro-pyrido-pyrimidinecompounds of the formula (I) and/or pharmaceutically acceptable saltsand/or solvates thereof,

wherein

Y is selected from O or NR³;

R¹ is selected from phenyl, pyridyl, pyrimidinyl, pyrazinyl,pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl,

or

-   -   —C(O)—R⁴        -   wherein        -   R⁴ is selected from C₁-C₈-alkyl, halo-C₁-C₈-alkyl,            hydroxy-C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl,            C₁-C₈-alkyl-sulfonyl-C₁-C₈-alkyl, heterocyclyl,            heterocyclyl-oxy, heterocyclyl-C₁-C₈-alkyl,            C₃-C₁₂-cycloalkyl, C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl,            heteroaryl, heteroaryl-oxy, heteroaryl-C₁-C₈-alkyl, hydroxy,            C₁-C₈-alkoxy, amino, N—C₁-C₈-alkyl-amino or            N,N-di-C₁-C₈-alkyl-amino,        -   wherein ‘C₁-C₈-alkyl’ in N—C₁-C₈-alkyl-amino and            N,N-di-C₁-C₈-alkyl-amino may be unsubstituted or substituted            by halogen, hydroxy or C₁-C₄-alkoxy;        -   wherein ‘C₃-C₁₂-cycloalkyl’ in C₃-C₁₂-cycloalkyl and            C₃-C₁₂-cycloalkyl-C₁-C₈-alkyl may be unsubstituted or            substituted by 1-5 substituents independently selected from            oxo, halogen, C₁-C₈-alkyl, halo-C₁-C₈-alkyl,            hydroxy-C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,            C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,            N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,            halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl or            C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl;        -   wherein ‘heterocyclyl’ is selected from oxiranyl,            aziridinyl, oxetanyl, thiethanyl, acetitinyl, pyrrolidinyl,            tetrahydrofuranyl, tetrahydrothiophenyl, 2,3-dihydrofuranyl,            2,5-dihydrofuranyl, 2,3-dihydrothiophenyl, 1-pyrrolinyl,            2-pyrrolinyl, 3-pyrrolinyl, tetrahydropyranyl, piperidinyl,            tetrahydrothiopyranyl, morpholinyl, thiomorpholinyl,            piperazinyl, azepanyl, thiepanyl or oxepanyl; each of which            is unsubstituted or substituted by 1-5 substituents            independently selected from oxo, halogen, C₁-C₈-alkyl,            halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl, hydroxyl,            C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,            N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,            C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,            hydroxy-C₁-C₈-alkyl-carbonyl or            C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl;            -   wherein ‘heterocyclyl’ can be attached at a heteroatom                or a carbon atom and where the N and/or S heteroatoms                can also optionally be oxidized to various oxidation                states;            -   wherein ‘heteroaryl’ is selected from            -   furanyl, thiophenyl, pyrrolyl, imidazolyl, pyrazolyl,                thiazolyl, isothiazolyl, oxazolyl, isoxazolyl,                1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-oxadiazolyl,                1,3,4-oxadiazolyl, 1,2,5-thiadiazolyl,                1,2,4-thiadiazolyl, 1,2,3-thiadiazolyl,                1,3,4-thiadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl,                1,2,5-triazolyl, pyridyl, pyrimidinyl, pyrazinyl,                pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl or                1,3,5-triazinyl; each of which is unsubstituted or                substituted by 1-5 substituents independently selected                from halogen, C₁-C₈-alkyl, halo-C₁-C₈-alkyl,                hydroxy-C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,                C₁-C₈-alkoxy-C₁-C₈-alkyl, amino, N—C₁-C₈-alkyl-amino,                N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,                halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl                or C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl; wherein                ‘heteroaryl’ can be attached at a heteroatom or a carbon                atom and where the N and/or S heteroatoms can also                optionally be oxidized to various oxidation states;

R² is selected from phenyl, naphthyl, pyridyl, pyrimidinyl, pyrazinyl,pyridazinyl, quinolinyl or isoquinolinyl, each of which is unsubstitutedor substituted by 1-5 substituents independently selected from halogen,cyano, nitro, C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,hydroxyl, C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl;

R³ is selected from H, C₁-C₄-alkyl or halo-C₁-C₄-alkyl; and

m is selected from 0 or 1.

DESCRIPTION OF THE DRAWINGS

FIG. 1 discloses the X-ray Powder Diffraction Pattern of Example 1citrate salt

FIG. 2 discloses the X-ray Powder Diffraction Pattern of Example 1fumarate salt

FIG. 3 discloses the X-ray Powder Diffraction Pattern of Example 1napadisylate salt

FIG. 4 discloses the X-ray Powder Diffraction Pattern of Example 67phosphate salt

FIG. 5 discloses the: X-ray Powder Diffraction Pattern of Example 67 HClsalt

FIG. 6 discloses the X-ray Powder Diffraction Pattern of Example 67hippurate salt

FIG. 7 discloses the X-ray Powder Diffraction Pattern of Example 1anhydrous form

FIG. 8 discloses the X-ray Powder Diffraction Pattern of Example 1trihydrate

FIG. 9 discloses the X-ray Powder Diffraction Pattern of Example 67anhydrous form

DETAILED DESCRIPTION OF THE INVENTION

Any formula given herein is intended to represent hydrates, solvates,and polymorphs of such compounds, and mixtures thereof.

Unless otherwise specified, the term “compounds of the invention” refersto compounds of formula (I) and subformulae thereof, salts of thecompounds, hydrates or solvates of the compounds, salts of the compoundsas well as stereoisomers (including diastereoisomers and enantiomers),tautomers and isotopically labeled compounds (including deuteriumsubstitutions).

As used herein, the term “a”, “an”, “the” and similar terms used in thecontext of the present invention, especially in the context of theclaims, are to be construed to cover both the singular and plural unlessotherwise indicated herein or clearly contradicted by the context.

All methods described herein can be performed in any suitable orderunless otherwise indicated herein or otherwise clearly contradicted bycontext. The use of any and all examples, or exemplary language, e.g.“such as”, provided herein is intended merely to better illuminate theinvention and does not pose a limitation on the scope of the inventionotherwise claimed.

The invention may be more fully appreciated by reference to thefollowing description, including the following glossary of terms and theconcluding examples. As used herein, the terms “including”, “containing”and “comprising” are used herein in their open, non-limiting sense.Where compounds of formula I are mentioned, this is meant to includealso the tautomers and N-oxides of the compounds of formula I.Tautomers, such as tautomers between keto- and enol form, lactam- andlactim form, amid form and imidic acid form or enamine form and imineform, can be present for example in the R1 or R2 portion of compounds offormula I. The nitrogen atoms of the tetrahydro-pyrido-pyrimidine coreof the compounds of formula I as well as nitrogen containingheterocyclyl and heteroaryl residues can form N-oxides.

Where the plural form is used for compounds, salts, and the like, thisis taken to mean also a single compound, salt, or the like.

The general terms used hereinbefore and hereinafter preferably havewithin the context of this disclosure the following meanings, unlessotherwise indicated:

As used herein, the term “alkyl” refers to a fully saturated branched,including single or multiple branching, or unbranched hydrocarbon moietyhaving up to 20 carbon atoms. Unless otherwise provided, alkyl refers tohydrocarbon moieties having 1 to 16 carbon atoms, 1 to 10 carbon atoms,1 to 7 carbon atoms, or 1 to 4 carbon atoms. Representative examples ofalkyl include, but are not limited to, methyl, ethyl, n-propyl,iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl,isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl,2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl and the like.Typically, alkyl groups have 1-7, more preferably 1-4 carbons.

As used herein, the term “halo-alkyl” refers to an alkyl as definedherein, which is substituted by one or more halo groups as definedherein. The halo-alkyl can be mono-halo-alkyl, di-halo-alkyl orpoly-halo-alkyl including per-halo-alkyl. A mono-halo-alkyl can have oneiodo, bromo, chloro or fluoro within the alkyl group. Di-halo-alky andpoly-halo-alkyl groups can have two or more of the same halo atoms or acombination of different halo groups within the alkyl. Typically thepoly-halo-alkyl contains up to 12, or 10, or 8, or 6, or 4, or 3, or 2halo groups. Non-limiting examples of halo-alkyl include fluoro-methyl,di-fluoro-methyl, tri-fluoro-methyl, chloro-methyl, di-chloro-methyl,tri-chloro-methyl, penta-fluoro-ethyl, hepta-fluoro-propyl,di-fluoro-chloro-methyl, di-chloro-fluoro-methyl, di-fluoro-ethyl,di-fluoro-propyl, di-chloro-ethyl and dichloro-propyl. A per-halo-alkylrefers to an alkyl having all hydrogen atoms replaced with halo atoms.

As used herein, the term “heterocyclyl” or “heterocyclic” refers to a 3to 7 membered monocyclic or 7 to 10 membered saturated or partiallysaturated ring or ring system, which contains at least one heteroatomselected from N, O and S, where the N and S can also optionally beoxidized to various oxidation states. ‘Heterocyclyl’ can be attached ata heteroatom or a carbon atom. ‘Heterocyclyl’ can include fused orbridged rings as well as spirocyclic rings. Examples of heterocyclesinclude oxiranyl, aziridinyl, oxetanyl, thiethanyl, acetitinyl,pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl,2,3-dihydrofuranyl, 2,5-dihydrofuranyl, 2,3-dihydrothiophenyl,1-pyrrolinyl, 2-pyrrolinyl, 3-pyrrolinyl, tetrahydropyranyl,piperidinyl, tetrahydrothiopyranyl, morpholinyl thiomorpholinyl,piperazinyl, azepanyl, thiepanyl and oxepanyl.

As used herein, the term “heteroaryl” refers to a 4-, 5-, 6-, or7-membered monocyclic, 7-, 8-, 9-, 10-, 11-, or 12-membered bicyclic or10-, 11-, 12-, 13-, 14- or 15-membered tricyclic unsaturated ring orring system—carrying the highest possible number of conjugated doublebonds in the ring(s), which contains at least one heteroatom selectedfrom N, O and S, wherein the N and S can also optionally be oxidized tovarious oxidation states. ‘Heteroaryl’ can be attached at a heteroatomor a carbon atom. ‘Heteroaryl’ can include fused or bridged rings aswell as spirocyclic rings. Examples of heteroaryl include furanyl,thiophenyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl,oxazolyl, isoxazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl,1,2,3-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-thiadiazolyl,1,2,4-thiadiazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, pyridyl, pyrimidinyl,pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl and1,3,5-triazinyl.

As used herein, the term “cycloalkyl” refers to saturated or partiallyunsaturated monocyclic, bicyclic or tricyclic hydrocarbon groups of 3-12carbon atoms. Unless otherwise provided, cycloalkyl refers to cyclichydrocarbon groups having between 3 and 10 ring carbon atoms or between3 and 7 ring carbon atoms. Exemplary bicyclic hydrocarbon groups includeoctahydroindyl, decahydronaphthyl. Exemplary tricyclic hydrocarbonbicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl,6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl,bicyclo[2.2.2]octy. Exemplary tetracyclic hydrocarbon groups includeadamantyl.

As used herein, the term “oxy” refers to an —O— linking group.

As used herein, the term “carboxy” or “carboxyl” is —COOH.

As used herein, all substituents are written in a way to show the orderof functional groups (groups) they are composed of. The functionalgroups are defined herein above.

“Treatment” includes prophylactic (preventive) and therapeutic treatmentas well as the delay of progression of a disease or disorder.

“Combination” refers to either a fixed combination in one dosage unitform, or a kit of parts for the combined administration where a compoundof the formula (I) and a combination partner (e.g. an other drug asexplained below, also referred to as “therapeutic agent” or “co-agent”)may be administered independently at the same time or separately withintime intervals, especially where these time intervals allow that thecombination partners show a cooperative, e.g. synergistic effect. Theterms “co-administration” or “combined administration” or the like asutilized herein are meant to encompass administration of the selectedcombination partner to a single subject in need thereof, e.g. a patient,and are intended to include treatment regimens in which the agents arenot necessarily administered by the same route of administration or atthe same time. The term “pharmaceutical combination” as used hereinmeans a product that results from the mixing or combining of more thanone active ingredient and includes both fixed and non-fixed combinationsof the active ingredients. The term “fixed combination” means that theactive ingredients, e.g. a compound of formula (I) and a combinationpartner, are both administered to a patient simultaneously in the formof a single entity or dosage. The term “non-fixed combination” meansthat the active ingredients, e.g. a compound of formula (I) and acombination partner, are both administered to a patient as separateentities either simultaneously, concurrently or sequentially with nospecific time limits, wherein such administration providestherapeutically effective levels of the two compounds in the body of thepatient. The latter also applies to cocktail therapy, e.g. theadministration of three or more active ingredients.

Various embodiments of the invention are described herein. It will berecognized that features specified in each embodiment may be combinedwith other specified features to provide further embodiments.

The invention further relates to pharmaceutically acceptable prodrugs ofa compound of formula (I). Particularly, the present invention alsorelates to pro-drugs of a compound of formula I as defined herein thatconvert in vivo to the compound of formula I as such. Any reference to acompound of formula I is therefore to be understood as referring also tothe corresponding pro-drugs of the compound of formula I, as appropriateand expedient.

The invention further relates to pharmaceutically acceptable metabolitesof a compound of formula (I).

In one embodiment, the invention provides a compound of the formula (I)and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ia)

wherein R¹, R² and Y are as defined above.

In one embodiment, the invention provides a compound of the formula (I)and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ia′)

wherein R¹, R² and Y are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ib)

wherein R¹ and R² are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ib′)

wherein R¹ and R² are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ic)

wherein R¹ and R² are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ic′)

wherein R¹ and R² are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Id)

wherein R⁴ and R² are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Id′)

wherein R⁴ and R² are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ie)

wherein R⁴ and R² are as defined above.

In another embodiment, the invention provides a compound of the formula(I) and/or a pharmaceutically acceptable salt and/or a solvate thereof,selected from a compound of the formula (Ie′)

wherein R⁴ and R² are as defined above.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie) or (Ie′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein R² is selected from naphthyl, pyridyl or pyrimidinyl; each ofwhich is unsubstituted or substituted by 1-3 substituents independentlyselected from halogen, cyano, nitro, C₁-C₈-alkyl, halo-C₁-C₈-alkyl,hydroxy-C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl,amino, N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino,C₁-C₈-alkyl-carbonyl, halo-C₁-C₈-alkyl-carbonyl,hydroxy-C₁-C₈-alkyl-carbonyl or C₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie) or (Ie′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein R² is selected from 3-pyridyl or 5-pyrimidinyl; each of which issubstituted by 1-2 substituents independently selected from halogen,cyano, nitro, C₁-C₈-alkyl, halo-C₁-C₈-alkyl, hydroxy-C₁-C₈-alkyl,hydroxyl, C₁-C₈-alkoxy, C₁-C₈-alkoxy-C₁-C₈-alkyl, amino,N—C₁-C₈-alkyl-amino, N,N-di-C₁-C₈-alkyl-amino, C₁-C₈-alkyl-carbonyl,halo-C₁-C₈-alkyl-carbonyl, hydroxy-C₁-C₈-alkyl-carbonyl orC₁-C₈-alkoxy-C₁-C₈-alkyl-carbonyl, wherein one substituents is locatedin the para position relative to the point of connection of R² to thecore of the compound.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie) or (Ie′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein R² is selected from 3-pyridyl or 5-pyrimidinyl; each of which issubstituted by 1-2 substituents independently selected from halogen,cyano, C₁-C₄-alkyl, halo-C₁-C₄-alkyl, C₁-C₄-alkoxy or amino, wherein onesubstituents is located in the para position relative to the point ofconnection of R² to the core of the compound.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′) (Ic), (Ic′), (Id), (Id′), (Ie) or (Ie′)and/or a pharmaceutically acceptable salt and/or a solvate thereof,wherein R² is selected from 3-pyridyl or 5-pyrimidinyl; each of which issubstituted by 1-2 substituents independently selected from fluoro,chloro, cyano, methyl, trifluoromethyl, methoxy or amino, wherein onesubstituents is located in the para position relative to the point ofconnection of R² to the core of the compound.

In another embodiment, the invention provides a compound of the formulae(I), (Ia) or (Ia′) and/or a pharmaceutically acceptable salt and/or asolvate thereof, wherein R³ is H.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) and/or a pharmaceuticallyacceptable salt and/or a solvate thereof, wherein R¹ is selected fromphenyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1,2,3-triazinyl,1,2,4-triazinyl or 1,3,5-triazinyl.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) and/or a pharmaceuticallyacceptable salt and/or a solvate thereof, wherein R¹ is selected frompyridyl or pyrimidinyl.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) and/or a pharmaceuticallyacceptable salt and/or a solvate thereof, wherein R¹ is —C(O)—R⁴,wherein R⁴ is as defined above.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) and/or a pharmaceuticallyacceptable salt and/or a solvate thereof, wherein R¹ is —C(O)—R⁴,wherein R⁴ is as defined above.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) wherein R¹ is —C(O)—R⁴; ora compound of the formulae (Id), (Id′), (Ie) or (Ie′) and/or apharmaceutically acceptable salt and/or a solvate thereof, wherein

-   -   R⁴ is selected from C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl,        heterocyclyl, heterocyclyl-C₁-C₈-alkyl, C₃-C₁₂-cycloalkyl,        heteroaryl, C₁-C₈-alkoxy or N,N-di-C₁-C₈-alkyl-amino,    -   wherein ‘C₁-C₈-alkyl’ in N,N-di-C₁-C₈-alkyl-amino may be        unsubstituted or substituted by halogen, hydroxy or        C₁-C₄-alkoxy;    -   wherein ‘C₃-C₁₂-cycloalkyl’ in C₃-C₁₂-cycloalkyl may be        unsubstituted or substituted by 1-3 substituents independently        selected from oxo, halogen, C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,        C₁-C₈-alkyl-carbonyl;    -   wherein ‘heterocyclyl’ is selected from pyrrolidinyl,        tetrahydropyranyl, piperidinyl, tetrahydrothiopyranyl,        morpholinyl or piperazinyl; each of which is unsubstituted or        substituted by 1-3 substituents independently selected from oxo,        halogen, C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,        C₁-C₈-alkyl-carbonyl;    -   wherein ‘heterocyclyl’ can be attached at a heteroatom or a        carbon atom and where the N and/or S heteroatoms can also        optionally be oxidized to various oxidation states;    -   wherein ‘heteroaryl’ is selected from    -   furanyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, isoxazolyl,        1,3,4-oxadiazolyl, pyridyl, pyrazinyl; each of which is        unsubstituted or substituted by 1-3 substituents independently        selected from halogen, C₁-C₈-alkyl, hydroxyl, C₁-C₈-alkoxy,        C₁-C₈-alkyl-carbonyl;    -   wherein ‘heteroaryl’ can be attached at a heteroatom or a carbon        atom and where the N and/or S heteroatoms can also optionally be        oxidized to various oxidation states.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) wherein R¹—C(O)—R⁴; or acompound of the formulae (Id), (Id′), (Ie) or (Ie′) and/or apharmaceutically acceptable salt and/or a solvate thereof, wherein

-   -   R⁴ is selected from heterocyclyl, C₄-C₈-cycloalkyl or        heteroaryl;    -   wherein ‘C₃-C₁₂-cycloalkyl’ may be unsubstituted or substituted        by 1-3 substituents independently selected from fluoro,        C₁-C₄-alkyl, hydroxyl, C₁-C₄-alkoxy;    -   wherein ‘heterocyclyl’ is selected from pyrrolidinyl,        tetrahydropyranyl, piperidinyl, tetrahydrothiopyranyl,        morpholinyl or piperazinyl; each of which is unsubstituted or        substituted by 1-3 substituents independently selected from oxo,        halogen, C₁-C₄-alkyl, hydroxyl, C₁-C₄-alkyl-carbonyl;    -   wherein ‘heterocyclyl’ can be attached at a heteroatom or a        carbon atom and where the N and/or S heteroatoms can also        optionally be oxidized to various oxidation states;    -   wherein ‘heteroaryl’ is selected from    -   furanyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, isoxazolyl,        1,3,4-oxadiazolyl, pyridyl, pyrazinyl; each of which is        unsubstituted or substituted by 1-3 substituents independently        selected from C₁-C₄-alkyl, hydroxyl;    -   wherein ‘heteroaryl’ can be attached at a heteroatom or a carbon        atom and where the N and/or S heteroatoms can also optionally be        oxidized to various oxidation states.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) wherein R¹—C(O)—R⁴; or acompound of the formulae (Id), (Id′), (Ie) or (Ie′) and/or apharmaceutically acceptable salt and/or a solvate thereof, wherein

-   -   R⁴ is selected from heterocyclyl;    -   wherein ‘heterocyclyl’ is selected from pyrrolidinyl,        tetrahydropyranyl, piperidinyl, tetrahydrothiopyranyl,        morpholinyl or piperazinyl; each of which is unsubstituted or        substituted by 1-3 substituents independently selected from oxo,        halogen, C₁-C₄-alkyl, hydroxyl, C₁-C₄-alkyl-carbonyl;    -   wherein ‘heterocyclyl’ can be attached at a heteroatom or a        carbon atom and where the N and/or S heteroatoms can also        optionally be oxidized to various oxidation states.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) wherein R¹ is —C(O)—R⁴; ora compound of the formulae (Id), (Id′), (Ie) or (Ie′) and/or apharmaceutically acceptable salt and/or a solvate thereof, wherein

-   -   R⁴ is selected from C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl,        C₁-C₈-alkoxy or N,N-di-C₁-C₈-alkyl-amino,    -   wherein ‘C₁-C₈-alkyl’ in N,N-di-C₁-C₈-alkyl-amino may be        unsubstituted or substituted by halogen, hydroxy or        C₁-C₄-alkoxy.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) wherein R¹ is —C(O)—R⁴; ora compound of the formulae (Id), (Id′), (Ie) or (Ie′) and/or apharmaceutically acceptable salt and/or a solvate thereof, wherein R⁴ isselected from C₁-C₈-alkyl.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) wherein R¹ is —C(O)—R⁴; ora compound of the formulae (Id), (Id′), (Ie) or (Ie′) and/or apharmaceutically acceptable salt and/or a solvate thereof, wherein

-   -   R² is selected from 3-pyridyl or 5-pyrimidinyl; each of which is        substituted by 1-2 substituents independently selected from        fluoro, chloro, cyano, methyl, trifluoromethyl, methoxy or        amino, wherein one substituents is located in the para position        relative to the point of connection of R² to the core of the        compound and    -   R⁴ is selected from heterocyclyl;    -   wherein ‘heterocyclyl’ is selected from pyrrolidinyl,        tetrahydropyranyl, piperidinyl, tetrahydrothiopyranyl,        morpholinyl or piperazinyl; each of which is unsubstituted or        substituted by 1-3 substituents independently selected from oxo,        halogen, C₁-C₄-alkyl, hydroxyl, C₁-C₄-alkyl-carbonyl;    -   wherein ‘heterocyclyl’ can be attached at a heteroatom or a        carbon atom and where the N and/or S heteroatoms can also        optionally be oxidized to various oxidation states.

In another embodiment, the invention provides a compound of the formulae(I), (Ia), (Ia′), (Ib), (Ib′), (Ic) or (Ic′) wherein R¹ is —C(O)—R⁴; ora compound of the formulae (Id), (Id′), (Ie) or (Ie′) and/or apharmaceutically acceptable salt and/or a solvate thereof, wherein

-   -   R² is selected from 3-pyridyl or 5-pyrimidinyl; each of which is        substituted by 1-2 substituents independently selected from        fluoro, chloro, cyano, methyl, trifluoromethyl, methoxy or        amino, wherein one substituents is located in the para position        relative to the point of connection of R² to the core of the        compound and    -   R⁴ is selected from C₁-C₈-alkyl, C₁-C₈-alkoxy-C₁-C₈-alkyl,        C₁-C₈-alkoxy or N,N-di-C₁-C₈-alkyl-amino,    -   wherein ‘C₁-C₈-alkyl’ in N,N-di-C₁-C₈-alkyl-amino may be        unsubstituted or substituted by halogen, hydroxy or        C₁-C₄-alkoxy.

In another embodiment individual compounds according to the inventionare those listed in the Examples section below.

In another embodiment, the invention provides a compound of the formula(I), selected from

-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   {(S)-3-[6-(2,4-Dimethoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   {3-[6-(2,4-Dimethoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   2-Methoxy-5-{4-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Methoxy-5-{4-[1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   1-{(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-propan-1-one;-   1-{3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-propan-1-one;-   {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   {3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   2-Amino-5-{4-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Amino-5-{4-[1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   (S)-(3-(6-(5-Fluoro-6-methoxypyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (3-(6-(5-Fluoro-6-methoxypyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (S)-2-Methoxy-5-(4-(1-(2-methoxyacetyl)pyrrolidin-3-yloxy)-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)nicotinonitrile;-   2-Methoxy-5-(4-(1-(2-methoxyacetyl)pyrrolidin-3-yloxy)-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)nicotinonitrile;-   (S)-5-(4-(1-(Cyclopentanecarbonyl)pyrrolidin-3-yloxy)-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)-2-methoxynicotinonitrile;-   5-(4-(1-(Cyclopentanecarbonyl)pyrrolidin-3-yloxy)-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)-2-methoxynicotinonitrile;-   (2,4-Dimethyl-oxazol-5-yl)-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   (2,4-Dimethyl-oxazol-5-yl)-{3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   Furan-3-yl-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   Furan-3-yl-{3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   Furan-3-yl-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   Furan-3-yl-{3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-3H-imidazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-3H-imidazol-4-yl)-methanone;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2-methyl-oxazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2-methyl-oxazol-4-yl)-methanone;-   (3-Methoxy-cyclobutyl)-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   (3-Methoxy-cyclobutyl)-{3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   ({(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanone;-   ({3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanone;-   1-(4-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanone;-   1-(4-{3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanone;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(4-methyl-oxazol-5-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(4-methyl-oxazol-5-yl)-methanone;-   5-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-1H-pyridin-2-one;-   5-{3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-1H-pyridin-2-one;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone;-   {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanone;-   {3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanone;-   {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-5-yl-methanone;-   {3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-5-yl-methanone;-   {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2-methyl-oxazol-4-yl)-methanone;-   {3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2-methyl-oxazol-4-yl)-methanone;-   {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2,2-dimethyl-tetrahydro-pyran-4-yl)-methanone;-   {3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2,2-dimethyl-tetrahydro-pyran-4-yl)-methanone;-   {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2,4-dimethyl-oxazol-5-yl)-methanone;-   {3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2,4-dimethyl-oxazol-5-yl)-methanone;-   (4,4-Difluoro-cyclohexyl)-{(S)-3-[6-(5,6-dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   (4,4-Difluoro-cyclohexyl)-{3-[6-(5,6-dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   2-Methoxy-5-{4-[(S)-1-(2-tetrahydro-pyran-4-yl-acetyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Methoxy-5-{4-[1-(2-tetrahydro-pyran-4-yl-acetyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   5-{4-[(S)-1-(2,4-Dimethyl-oxazole-5-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-2-methoxy-nicotinonitrile;-   5-{4-[1-(2,4-Dimethyl-oxazole-5-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-2-methoxy-nicotinonitrile;-   5-{4-[(S)-1-(2,2-Dimethyl-tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-2-methoxy-nicotinonitrile;-   5-{4-[1-(2,2-Dimethyl-tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-2-methoxy-nicotinonitrile;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(5-methyl-oxazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(5-methyl-oxazol-4-yl)-methanone;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(5-methyl-isoxazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(5-methyl-isoxazol-4-yl)-methanone;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-isoxazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-isoxazol-4-yl)-methanone;-   {(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-isoxazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-isoxazol-4-yl)-methanone;-   Isoxazol-3-yl-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   Isoxazol-3-yl-{3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   Isoxazol-5-yl-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   Isoxazol-5-yl-{3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   2-Methoxy-5-{4-[(S)-1-(thiazole-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Methoxy-5-{4-[1-(thiazole-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Methoxy-5-{4-[(S)-1-(1-methyl-1H-pyrazole-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Methoxy-5-{4-[1-(1-methyl-1H-pyrazole-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Methoxy-5-{4-[(S)-1-(1-methyl-1H-pyrazole-3-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   2-Methoxy-5-{4-[1-(1-methyl-1H-pyrazole-3-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile;-   (2,2-Dimethyl-tetrahydro-pyran-4yl)-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   (2,2-Dimethyl-tetrahydro-pyran-4yl)-{3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   (1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone;-   (S)-(2,4-Dimethyloxazol-5-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   (2,4-Dimethyloxazol-5-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(thiazol-5-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(thiazol-5-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(1-methyl-1H-pyrazol-5-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(1-methyl-1H-pyrazol-5-yl)methanone;-   4-((S)-3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carbonyl)pyrrolidin-2-one;-   4-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carbonyl)pyrrolidin-2-one;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(pyridin-3-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(pyridin-3-yl)methanone;-   (S)-(1H-Imidazol-4-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   (1H-Imidazol-4-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   5-((S)-3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carbonyl)pyrrolidin-2-one;-   5-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carbonyl)pyrrolidin-2-one;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(pyridin-4-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(pyridin-4-yl)methanone;-   (S)-(1,3-Dimethyl-1H-pyrazol-4-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   (1,3-Dimethyl-1H-pyrazol-4-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(1H-pyrazol-4-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(1H-pyrazol-4-yl)methanone;-   (S)-(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(5-methyl-1,3,4-oxadiazol-2-yl)methanone;-   (3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(5-methyl-1,3,4-oxadiazol-2-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(pyrazin-2-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(pyrazin-2-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(1-methyl-1H-imidazol-4-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(1-methyl-1H-imidazol-4-yl)methanone;-   {(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-pyrazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-pyrazol-4-yl)-methanone;-   {(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-thiazol-4-yl-methanone;-   {3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-thiazol-4-yl-methanone;-   {(S)-3-[6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   {3-[6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   (S)-(3-(6-(6-Amino-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (3-(6-(6-Amino-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)azetidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   {(S)-3-[6-(2-Methoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   {3-[6-(2-Methoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;-   [(S)-3-(6-Quinolin-3-yl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-(tetrahydro-pyran-4-yl)-methanone;-   [3-(6-Quinolin-3-yl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-(tetrahydro-pyran-4-yl)-methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (S)-1-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)-3,3-dimethylbutan-1-one;-   1-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)-3,3-dimethylbutan-1-one;-   1-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-propan-1-one;-   1-{3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-propan-1-one;-   2-Methoxy-5-[4-((S)-1-propionyl-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]-nicotinonitrile;-   2-Methoxy-5-[4-(1-propionyl-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]-nicotinonitrile;-   (S)-6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-4-(1-(pyridin-2-yl)    pyrrolidin-3-yloxy)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidine;-   6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-4-(1-(pyridin-2-yl)    pyrrolidin-3-yloxy)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidine;-   (S)-6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-4-(1-(pyrimidin-2-yl)pyrrolidin-3-yloxy)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidine;-   6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-4-(1-(pyrimidin-2-yl)pyrrolidin-3-yloxy)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidine;-   (S)-1-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)propan-1-one;-   1-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)propan-1-one;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (S)-2-Methoxy-5-(4-(1-(tetrahydro-2H-pyran-4-carbonyl)pyrrolidin-3-ylamino)-7,    8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)nicotinonitrile;-   2-Methoxy-5-(4-(1-(tetrahydro-2H-pyran-4-carbonyl)pyrrolidin-3-ylamino)-7,    8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)nicotinonitrile;-   (S)-1-(4-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidine-1-carbonyl)    piperidin-1-yl)ethanone;-   1-(4-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidine-1-carbonyl)    piperidin-1-yl)ethanone;-   (2,2-Dimethyltetrahydro-2H-pyran-4-yl)    ((S)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   (2,2-Dimethyltetrahydro-2H-pyran-4-yl)    (3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(oxazol-5-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(oxazol-5-yl)methanone;-   ((S)-3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)((1s,4R)-4-methoxycyclohexyl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)((1s,4R)-4-methoxycyclohexyl)methanone;-   ((S)-3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)((1r,4S)-4-methoxycyclohexyl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)((1r,4S)-4-methoxycyclohexyl)methanone;-   ((1s,4R)-4-Hydroxycyclohexyl)((S)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   ((1s,4R)-4-Hydroxycyclohexyl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   ((1r,4S)-4-Hydroxycyclohexyl)((S)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   ((1r,4S)-4-Hydroxycyclohexyl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(1-methyl-1H-imidazol-4-yl)methanone;-   (3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(1-methyl-1H-imidazol-4-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(oxazol-5-yl)methanone;-   (3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(oxazol-5-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(oxazol-4-yl)methanone;-   (3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(oxazol-4-yl)methanone;-   (2,2-Dimethyltetrahydro-2H-pyran-4-yl)((S)-3-(6-(6-methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   (2,2-Dimethyltetrahydro-2H-pyran-4-yl)(3-(6-(6-methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone;-   (S)-1-(3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)propan-1-one;-   1-(3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)propan-1-one;-   (S)-(3-(6-(5-Chloro-6-methoxypyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (3-(6-(5-Chloro-6-methoxypyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (3-(6-(6-Methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)(tetrahydro-2H-pyran-4-yl)methanone;-   (Tetrahydro-pyran-4-yl)-{(S)-3-{6-(5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl}-methanone;-   (Tetrahydro-pyran-4-yl)-{3-{6-(5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl}-methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(4-methylpiperazin-1-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(4-methylpiperazin-1-yl)methanone;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(morpholino)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(morpholino)methanone;-   (S)-(4-Hydroxypiperidin-1-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   4-Hydroxypiperidin-1-yl)(3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)methanone;-   (S)—N-(2-Hydroxyethyl)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)-N-methylpyrrolidine-1-carboxamide;-   N-(2-Hydroxyethyl)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)-N-methylpyrrolidine-1-carboxamide;-   (S)-1-(4-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carbonyl)piperazin-1-yl)ethanone;-   1-(4-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carbonyl)piperazin-1-yl)ethanone;-   (S)-2-Methoxy-5-(4-(1-(morpholine-4-carbonyl)pyrrolidin-3-yloxy)-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)nicotinonitrile;-   2-Methoxy-5-(4-(1-(morpholine-4-carbonyl)pyrrolidin-3-yloxy)-7,8-dihydropyrido[4,3-d]pyrimidin-6(5H)-yl)nicotinonitrile;-   (S)-(3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(oxazol-4-yl)methanone;-   (3-(6-(6-Methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl)(oxazol-4-yl)methanone;-   1-(4-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanone;-   1-(4-{3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanone;-   {(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-3H-imidazol-4-yl)-methanone;-   {3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-3H-imidazol-4-yl)-methanone;-   {(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-5-yl-methanone;-   {3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-5-yl-methanone;-   {(S)-3-[6-(6-Methoxy-pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone;    or-   {3-[6-(6-Methoxy-pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone.

Compounds of the formula (I) may have different isomeric forms. As usedherein, the term “an optical isomer” or “a stereoisomer” refers to anyof the various stereo isomeric configurations which may exist for agiven compound of the present invention and includes geometric isomers.It is understood that a substituent may be attached at a chiral centerof a carbon atom. Therefore, the invention includes enantiomers,diastereomers or racemates of the compound. “Enantiomers” are a pair ofstereoisomers that are non-superimposable mirror images of each other. A1:1 mixture of a pair of enantiomers is a “racemic” mixture. The term isused to designate a racemic mixture where appropriate.“Diastereoisomers” are stereoisomers that have at least two asymmetricatoms, but which are not mirror-images of each other. The absolutestereochemistry is specified according to the Cahn-Ingold-Prelog R—Ssystem. When a compound is a pure enantiomer the stereochemistry at eachchiral carbon may be specified by either R or S. Resolved compoundswhose absolute configuration is unknown can be designated (+) or (−)depending on the direction (dextro- or levorotatory) which they rotateplane polarized light at the wavelength of the sodium D line. Certain ofthe compounds described herein contain one or more asymmetric centers oraxes and may thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)— or (S)—. The present invention is meant toinclude all such possible isomers, including racemic mixtures, opticallypure forms and intermediate mixtures. Optically active (R)- and(S)-isomers may be prepared using chiral synthons or chiral reagents, orresolved using conventional techniques. If the compound contains adouble bond, the substituent may be E or Z configuration. If thecompound contains a disubstituted cycloalkyl, the cycloalkyl substituentmay have a cis- or trans-configuration. All tautomeric forms are alsointended to be included.

As used herein, the term “pharmaceutically acceptable salts” refers tosalts that retain the biological effectiveness and properties of thecompounds of this invention and, which typically are not biologically orotherwise undesirable. In many cases, the compounds of the presentinvention are capable of forming acid and/or base salts by virtue of thepresence of amino and/or carboxyl groups or groups similar thereto.

Pharmaceutically acceptable acid addition salts can be formed withinorganic acids and organic acids, e.g., acetate, aspartate, benzoate,besylate, bromide/hydrobromide, bicarbonate/carbonate,bisulfate/sulfate, camphorsulfornate, chloride/hydrochloride,chlortheophyllonate, citrate, ethandisulfonate, fumarate, gluceptate,gluconate, glucuronate, hippurate, hydroiodide/iodide, isethionate,lactate, lactobionate, laurylsulfate, malate, maleate, malonate,mandelate, mesylate, methylsulphate, naphthoate, napsylate, nicotinate,nitrate, octadecanoate, oleate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate,propionate, stearate, succinate, sulfosalicylate, tartrate, tosylate andtrifluoroacetate salts.

Inorganic acids from which salts can be derived include, for example,hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, and the like.

Organic acids from which salts can be derived include, for example,acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid,malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid,benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,toluenesulfonic acid, sulfosalicylic acid, and the like.Pharmaceutically acceptable base addition salts can be formed withinorganic and organic bases.

Inorganic bases from which salts can be derived include, for example,ammonium salts and metals from columns 1 to 12 of the periodic table. Incertain embodiments, the salts are derived from sodium, potassium,ammonium, calcium, magnesium, iron, silver, zinc, and copper;particularly suitable salts include ammonium, potassium, sodium, calciumand magnesium salts.

Organic bases from which salts can be derived include, for example,primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines, basic ionexchange resins, and the like. Certain organic amines includeisopropylamine, benzathine, cholinate, diethanolamine, diethylamine,lysine, meglumine, piperazine and tromethamine.

The pharmaceutically acceptable salts of the present invention can besynthesized from a parent compound, a basic or acidic moiety, byconventional chemical methods. Generally, such salts can be prepared byreacting free acid forms of these compounds with a stoichiometric amountof the appropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate,bicarbonate or the like), or by reacting free base forms of thesecompounds with a stoichiometric amount of the appropriate acid. Suchreactions are typically carried out in water or in an organic solvent,or in a mixture of the two. Generally, use of non-aqueous media likeether, ethyl acetate, ethanol, isopropanol, or acetonitrile isdesirable, where practicable. Lists of additional suitable salts can befound, e.g., in “Remington's Pharmaceutical Sciences”, 20th ed., MackPublishing Company, Easton, Pa., (1985); and in “Handbook ofPharmaceutical Salts: Properties, Selection, and Use” by Stahl andWermuth (Wiley-VCH, Weinheim, Germany, 2002).

For isolation or purification purposes it is also possible to usepharmaceutically unacceptable salts, for example picrates orperchlorates. For therapeutic use, only pharmaceutically acceptablesalts or free compounds are employed.

In view of the close relationship between the novel compounds of theformula (I) in free form and those in the form of their salts, includingthose salts that can be used as intermediates, for example in thepurification or identification of the novel compounds, any reference tothe compounds or a compound of the formula (I) hereinbefore andhereinafter is to be understood as referring to the compound in freeform and/or also to one or more salts thereof, as appropriate andexpedient, as well as to one or more solvates, e.g. hydrates.

Any formula given herein is also intended to represent unlabeled formsas well as isotopically labeled forms of the compounds. Isotopicallylabeled compounds have structures depicted by the formulas given hereinexcept that one or more atoms are replaced by an atom having a selectedatomic mass or mass number. Examples of isotopes that can beincorporated into compounds of the invention include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, and chlorine,such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸F ³¹P, ³²P, ³⁵S, ³⁶Cl, ¹²⁵Irespectively. The invention includes various isotopically labeledcompounds as defined herein, for example those into which radioactiveisotopes, such as ³H and ¹⁴C or those into which non-radioactiveisotopes, such as ²H and ¹³C, are present. Such isotopically labelledcompounds are useful in metabolic studies (with ¹⁴C), reaction kineticstudies (with, for example ²H or ³H), detection or imaging techniques,such as positron emission tomography (PET) or single-photon emissioncomputed tomography (SPECT) including drug or substrate tissuedistribution assays, or in radioactive treatment of patients. Inparticular, an ¹⁸F or labeled compound may be particularly desirable forPET or SPECT studies. Isotopically labeled compounds of this inventionand prodrugs thereof can generally be prepared by carrying out theprocedures disclosed in the schemes or in the examples and preparationsdescribed below by substituting a readily available isotopically labeledreagent for a non-isotopically labeled reagent.

Further, substitution with heavier isotopes, particularly deuterium(i.e., ²H or D) may afford certain therapeutic advantages resulting fromgreater metabolic stability, for example increased in vivo half-life orreduced dosage requirements or an improvement in therapeutic index. Itis understood that deuterium in this context is regarded as asubstituent of a compound of the formula (I). The concentration of sucha heavier isotope, specifically deuterium, may be defined by theisotopic enrichment factor. The term “isotopic enrichment factor” asused herein means the ratio between the isotopic abundance and thenatural abundance of a specified isotope. If a substituent in a compoundof this invention is denoted deuterium, such compound has an isotopicenrichment factor for each designated deuterium atom of at least 3500(52.5% deuterium incorporation at each designated deuterium atom), atleast 4000 (60% deuterium incorporation), at least 4500 (67.5% deuteriumincorporation), at least 5000 (75% deuterium incorporation), at least5500 (82.5% deuterium incorporation), at least 6000 (90% deuteriumincorporation), at least 6333.3 (95% deuterium incorporation), at least6466.7 (97% deuterium incorporation), at least 6600 (99% deuteriumincorporation), or at least 6633.3 (99.5% deuterium incorporation).

Isotopically-labeled compounds of the formula (I) can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described in the accompanying Examplesand Preparations using an appropriate isotopically-labeled reagents inplace of the non-labeled reagent previously employed.

Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d₆-acetone, d₆-DMSO.

Compounds of the invention, i.e. compounds of the formula (I) thatcontain groups capable of acting as donors and/or acceptors for hydrogenbonds may be capable of forming co-crystals with suitable co-crystalformers. These co-crystals may be prepared from compounds of the formula(I) by known co-crystal forming procedures. Such procedures includegrinding, heating, co-subliming, co-melting, or contacting in solutioncompounds of the formula (I) with the co-crystal former undercrystallization conditions and isolating co-crystals thereby formed.Suitable co-crystal formers include those described in WO 2004/078163.Hence the invention further provides co-crystals comprising a compoundof the formula (I).

Any asymmetric atom (e.g., carbon or the like) of the compound(s) of thepresent invention can be present in racemic or enantiomericallyenriched, for example the (R)-, (S)- or (R,S)-configuration. In certainembodiments, each asymmetric atom has at least 50% enantiomeric excess,at least 60% enantiomeric excess, at least 70% enantiomeric excess, atleast 80% enantiomeric excess, at least 90% enantiomeric excess, atleast 95% enantiomeric excess, or at least 99% enantiomeric excess inthe (R)- or (S)-configuration. Substituents at atoms with unsaturatedbonds may, if possible, be present in cis-(Z)- or trans-(E)-form.

Accordingly, as used herein a compound of the present invention can bein the form of one of the possible isomers, rotamers, atropisomers,tautomers or mixtures thereof, for example, as substantially puregeometric (cis or trans) isomers, diastereomers, optical isomers(antipodes), racemates or mixtures thereof.

Mixtures of isomers obtainable according to the invention can beseparated in a manner known to those skilled in the art into theindividual isomers; diastereoisomers can be separated, for example, bypartitioning between polyphasic solvent mixtures, recrystallisationand/or chromatographic separation, for example over silica gel or bye.g. medium pressure liquid chromatography over a reversed phase column,and racemates can be separated, for example, by the formation of saltswith optically pure salt-forming reagents and separation of the mixtureof diastereoisomers so obtainable, for example by means of fractionalcrystallisation, or by chromatography over optically active columnmaterials.

Any resulting racemates of final products or intermediates can beresolved into the optical antipodes by known methods, e.g., byseparation of the diastereomeric salts thereof, obtained with anoptically active acid or base, and liberating the optically activeacidic or basic compound. In particular, a basic moiety may thus beemployed to resolve the compounds of the present invention into theiroptical antipodes, e.g., by fractional crystallization of a salt formedwith an optically active acid, e.g., tartaric acid, dibenzoyl tartaricacid, diacetyl tartaric acid, di-O,O′-p-toluoyl tartaric acid, mandelicacid, malic acid or camphor-10-sulfonic acid. Racemic products can alsobe resolved by chiral chromatography, e.g., high pressure liquidchromatography (HPLC) using a chiral adsorbent. Compounds of the presentinvention are either obtained in the free form, as a salt thereof, or asprodrug derivatives thereof.

When both a basic group and an acid group are present in the samemolecule, the compounds of the present invention may also form internalsalts, e.g., zwitterionic molecules.

The present invention also provides pro-drugs of the compounds of thepresent invention that converts in vivo to the compounds of the presentinvention. A pro-drug is an active or inactive compound that is modifiedchemically through in vivo physiological action, such as hydrolysis,metabolism and the like, into a compound of this invention followingadministration of the prodrug to a subject. The suitability andtechniques involved in making and using pro-drugs are well known bythose skilled in the art. Prodrugs can be conceptually divided into twonon-exclusive categories, bioprecursor prodrugs and carrier prodrugs.See The Practice of Medicinal Chemistry, Ch. 31-32 (Ed. Wermuth,Academic Press, San Diego, Calif., 2001). Generally, bioprecursorprodrugs are compounds, which are inactive or have low activity comparedto the corresponding active drug compound, which contain one or moreprotective groups and are converted to an active form by metabolism orsolvolysis. Both the active drug form and any released metabolicproducts should have acceptably low toxicity.

Carrier prodrugs are drug compounds that contain a transport moiety,e.g. that improve uptake and/or localized delivery to a site(s) ofaction. Desirably for such a carrier prodrug, the linkage between thedrug moiety and the transport moiety is a covalent bond, the prodrug isinactive or less active than the drug compound, and any releasedtransport moiety is acceptably non-toxic. For prodrugs where thetransport moiety is intended to enhance uptake, typically the release ofthe transport moiety should be rapid. In other cases, it is desirable toutilize a moiety that provides slow release, e.g., certain polymers orother moieties, such as cyclodextrins. Carrier prodrugs can, forexample, be used to improve one or more of the following properties:increased lipophilicity, increased duration of pharmacological effects,increased site-specificity, decreased toxicity and adverse reactions,and/or improvement in drug formulation (e.g., stability, watersolubility, suppression of an undesirable organoleptic or physiochemicalproperty). For example, lipophilicity can be increased by esterificationof (a) hydroxyl groups with lipophilic carboxylic acids (e.g., acarboxylic acid having at least one lipophilic moiety), or (b)carboxylic acid groups with lipophilic alcohols (e.g., an alcohol havingat least one lipophilic moiety, for example aliphatic alcohols).

Exemplary prodrugs are, e.g., esters of free carboxylic acids and S-acylderivatives of thiols and O-acyl derivatives of alcohols or phenols,wherein acyl has a meaning as defined herein. Suitable prodrugs areoften pharmaceutically acceptable ester derivatives convertible bysolvolysis under physiological conditions to the parent carboxylic acid,e.g., lower alkyl esters, cycloalkyl esters, lower alkenyl esters,benzyl esters, mono- or di-substituted lower alkyl esters, such as theomega-(amino, mono- or di-lower alkylamino, carboxy, loweralkoxycarbonyl)-lower alkyl esters, the alpha-(lower alkanoyloxy, loweralkoxycarbonyl or di-lower alkylaminocarbonyl)-lower alkyl esters, suchas the pivaloyloxymethyl ester and the like conventionally used in theart. In addition, amines have been masked as arylcarbonyloxymethylsubstituted derivatives which are cleaved by esterases in vivo releasingthe free drug and formaldehyde (Bundgaard, J. Med. Chem. 2503 (1989)).

Moreover, drugs containing an acidic NH group, such as imidazole, imide,indole and the like, have been masked with N-acyloxymethyl groups(Bundgaard, Design of Prodrugs, Elsevier (1985)). Hydroxy groups havebeen masked as esters and ethers. EP 039,051 (Sloan and Little)discloses Mannich-base hydroxamic acid prodrugs, their preparation anduse.

Furthermore, the compounds of the present invention, including theirsalts, can also be obtained in the form of their hydrates, or includeother solvents used for their crystallization. The compounds of thepresent invention may inherently or by design form solvates withpharmaceutically acceptable solvents (including water); therefore, it isintended that the invention embrace both solvated and unsolvated forms.The term “solvate” refers to a molecular complex of a compound of thepresent invention (including pharmaceutically acceptable salts thereof)with one or more solvent molecules. Such solvent molecules are thosecommonly used in the pharmaceutical art, which are known to be innocuousto the recipient, e.g., water, ethanol, and the like. The term “hydrate”refers to the complex where the solvent molecule is water.

The compounds of the present invention, including salts, hydrates andsolvates thereof, may inherently or by design form polymorphs.

The invention relates in a second aspect to the manufacture of acompound of formula I. The compounds of formula I or salts thereof areprepared in accordance with processes known per se, though notpreviously described for the manufacture of the compounds of the formulaI.

General Reaction Processes:

In one embodiment, the invention relates to a process for manufacturinga compound of formula I (Method A) comprising steps a, d, e, b, and c.

The compound of formula I is obtained via the step b of deprotecting PG²from the compound of formula (E), wherein PG² represents a suitableprotecting group, preferable a benzyl group, and the other substituentsare as defined above,

followed by reaction step c with

R²-Hal,

Wherein R² is defined above and Hal represents halogen, particularlyiodo or bromo; under customary Buchwald-Hartwig conditions using aligand such as X-Phos, di-tert-butyl(2′-methylbiphenyl-2-yl)phosphine or2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl with a palladiumcatalyst such as Pd₂(dba)₃ or Pd₂(dba)₃.CHCl₃ or Pd(OAc)₂, preferablyPd₂(dba)₃ with X-Phos, in the presence of a base such as preferablyCs₂CO₃ or tert-BuONa, in an organic solvent such as an ether, preferablydioxane or THF. The reaction is preferably stirred at a temperature ofapproximately 80-120° C., preferably 120° C. The reaction is preferablycarried out under an inert gas such as nitrogen or argon.

The compound of formula (E) is prepared comprising the step d ofdeprotecting PG¹ from the compound of formula (C), wherein PG¹represents a suitable protecting group, for example BOC, and the othersubstituents are as defined above,

followed by coupling reaction step e with

R¹-Act,

step e1: Where R¹ is —C(O)—R⁴, wherein R⁴ is defined above, and Actrepresents an activating group or a hydroxy group: The coupling reactionis an amide, urea or carbamic ester formation. There are many known waysof preparing amides urea or carbamic ester. The coupling reaction stepcan be carried out with Act representing an activating group, preferablyin a one step procedure or with Act representing a hydroxy group eitherinvolving a one or two step procedure. For examples of amide bondformations, see Mantalbetti, C. A. G. N and Falque, V., Amide bondformation and peptide coupling, Tetrahedron, 2005, 61(46), pp10827-10852 and references cited therein. For examples of ureasynthesis, see Sartori, G.; Maggi, R. Acyclic and cyclic ureas, Scienceof Synthesis (2005), 18, 665-758; Gallou, Isabelle. Unsymmetrical ureasSynthetic methodologies and application in drug design, OrganicPreparations and Procedures International (2007), 39(4), 355-383. Forexamples of carbamate synthesis see Adams, Philip; Baron, Frank A.Esters of carbamic acid, Chemical Reviews (1965), 65(5), 567-602. Theexamples provided herein are thus not intended to be exhaustive, butmerely illustrative;

step e2: Where R¹ is selected from phenyl, pyridyl, pyrimidinyl,pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl or1,3,5-triazinyl and Act represents halogen, particularly iodo or bromo:The coupling reaction is carried out in the presence of an amine basesuch as N,N-diisopropylethylamine. The reaction is carried out in thepresence of an organic solvent or, preferably without a solvent undermicrowave heating e.g. at 160° C. for 20 min. Alternatively, thereaction can be carried out under customary Buchwald-Hartwig conditionsusing a ligand such as X-Phos or2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl with a palladiumcatalyst such as Pd₂(dba)₃ or Pd₂(dba)₃.CHCl₃ or Pd(OAc)₂, preferablyPd₂(dba)₃ with X-Phos, in the presence of a base such as preferablyCs₂CO₃ or tert-BuONa, in an organic solvent such as an ether, preferablydioxane or THF. The reaction is preferably stirred at a temperature ofapproximately 80-120° C., preferably 120° C. The reaction is preferablycarried out under an inert gas such as nitrogen or argon.

The compound of formula (C) is prepared comprising the step a ofcoupling a compound of formula (A), wherein PG² represents a suitableprotecting group, for example a benzyl group and X represents halogen,particularly chloro, or hydroxy; with a compound of formula (B) whereinPG¹ represents a suitable protecting group, for example BOC, and theother substituents are as defined above,

step a1: Where YH is OH and X represents halogen: The reaction takesplace in the presence of a suitable base such as sodium hydroxide (NaH)or potassium t-butoxide (tBuOK) and polar organic solvent such as THF,2-methyltetrahydrofuran or Dioxane under inert gas conditions at roomtemperature.

step a2: Where YH is NR³H and X represents halogen: The reaction takesplace in the presence of a suitable base such as for example potassiumcarbonate or a suitable amine base such as triethylamine orN,N-diisopropylethylamine at elevated temperature (e.g. 120° C.) for20-48 h. Typical conditions comprise the use of 1.0 equivalent of acompound of formula (A), 1.0 equivalent of a compound of formula (B) and1.5 equivalents of the base at 120° C. for 48 h.

step a3: Where YH is NR³H and X represents hydroxy: A base promotedphosphonium coupling reaction is employed, whereby a compound of theformula (A) in a suitable solvent such as acetonitrile is reacted with aphosphonium salt such asbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate(BOP) in the presence of a base such as1,8-diaza-7-bicyclo[5.4.0]undecene (DBU) followed by addition of acompound of the formula (B). The reaction mixture is preferably stirredat a temperature of 20° C. to 90° C. for 18-72 h. The reaction maypreferably be carried out under an inert gas, e.g. nitrogen or argon.Typical conditions comprise of 1 equivalent of a compound of the formula(A), 1.0-1.5 equivalents of BOP, 2.0-4.0 equivalents of DBU and 2.0-3.0equivalents of a compound of the formula (B) in acetonitrile at 65° C.for 72 hours under argon.

In another embodiment, the invention relates to a process formanufacturing a compound of formula I, comprising steps a, d and e asdefined above, starting from a compound of formula (A) wherein PG²represents R² (Method A-a).

The process for manufacturing a compound of formula (A) wherein PG²represents R² comprises the steps f, g, h, i and optionally k.

step f) Quaternarization of compound of formula (F), R⁸=alkyl e.g.benzyl with compound of general formula R⁹—Hal, wherein R⁹ representsalkyl, particularly methyl and Hal represents halogen, particularly iodoor bromo, under customary condition using in particular acetone asorganic solvent leads to the formation of compound of general formula(G).

step g) The compound of formula (H) is prepared by reaction of R²—NH₂with compound of general formula (G). The reaction is performed by usinga base such a in particular K₂CO₃ in an organic solvent such as inparticular a 2/1 mixture of ethanol and water and heating the reactionmixture at 80-100° C., in particular 80° C.

step h) The compound of formula (J) is prepared by reaction of acompound of formula (H) with base such as in particular NaH and compoundof general formula (R¹⁰O)₂CO, wherein R¹⁰ represents alkyl, particularlymethyl. The reaction mixture is stirred under high temperature (90° C.).

step i) The compound of formula (A), X=OH is prepared by pyrimidine ringformation, reacting the compound of formula (J) with formamidine acetatein the presence of a base such as sodium methoxide and in an organicsolvent such as methanol at elevated temperature such as 900° C. for2-18 h.

step k) The compound of formula (A), X=Cl is prepared by reaction of acompound of formula (A), X=OH with phosphoryl chloride in the presenceof a base such as triethylamine in an organic solvent such as toluene atelevated temperature such as 100° C. for 12-18 h.

In another embodiment, the invention relates to a process formanufacturing a compound of formula I (Method B) comprising steps a, b,c, d, and e.

The compound of formula I is obtained via the step d of deprotecting PG¹from the compound of formula, (D), wherein PG¹ represents a suitableprotecting group, preferable a BOC group, and the other substituents areas defined above,

followed by reaction step e with

R¹-Act,

step e1: Where R¹ is —C(O)—R⁴, wherein R⁴ is defined above, and Actrepresents an activating group or a hydroxy group: The coupling reactionis an amide, urea or carbamic ester formation. There are many known waysof preparing amides urea or carbamic ester. The coupling reaction stepcan be carried out with Act representing an activating group, preferablyin a one step procedure or with Act representing a hydroxy group eitherinvolving a one or two step procedure. For examples of amide bondformations, see Mantalbetti, C. A. G. N and Falque, V., Amide bondformation and peptide coupling, Tetrahedron, 2005, 61(46), pp10827-10852 and references cited therein. For examples of ureasynthesis, see Sartori, G.; Maggi, R. Acyclic and cyclic ureas, Scienceof Synthesis (2005), 18, 665-758; Gallou, Isabelle. Unsymmetrical ureasSynthetic methodologies and application in drug design, OrganicPreparations and Procedures International (2007), 39(4), 355-383. Forexamples of carbamate synthesis see Adams, Philip; Baron, Frank A.Esters of carbamic acid, Chemical Reviews (1965), 65(5), 567-602. Theexamples provided herein are thus not intended to be exhaustive, butmerely illustrative;

step e2: Where R¹ is selected from phenyl, pyridyl, pyrimidinyl,pyrazinyl, pyridazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl or1,3,5-triazinyl and Act represents halogen, particularly iodo or bromo:The coupling reaction is carried out in the presence of an amine basesuch as N,N-diisopropylethylamine. The reaction is carried out in thepresence of an organic solvent or, preferably without a solvent undermicrowave heating e.g. at 160° C. for 20 min. Alternatively, thereaction can be carried out under customary Buchwald-Hartwig conditionsusing such a ligand such as X-Phos or2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl with a palladiumcatalyst such as Pd₂(dba)₃ or Pd₂(dba)₃.CHCl₃ or Pd(OAc)₂, preferablyPd₂(dba)₃ with X-Phos, in the presence of a base such as preferablyCs₂CO₃ or tert-BuONa, in an organic solvent such as an ether, preferablydioxane or THF. The reaction is preferably stirred at a temperature ofapproximately 80-120° C., preferably 120° C. The reaction is preferablycarried out under an inert gas such as nitrogen or argon.

The compound of formula (D) is prepared comprising the step b ofdeprotecting PG² from the compound of formula (C), wherein PG²represents a suitable protecting group, for example a benzyl group, andthe other substituents are as defined above,

followed by reaction step c with

R²-Hal,

Wherein R² is defined above and Hal represents halogen, particularlyiodo or bromo; under customary Buchwald-Hartwig conditions using such aligand such as X-Phos or2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl with a palladiumcatalyst such as Pd₂(dba)₃ or Pd₂(dba)₃.CHCl₃ or Pd(OAc)₂, preferablyPd₂(dba)₃ with X-Phos, in the presence of a base such as preferablyCs₂CO₃ or tert-BuONa, in an organic solvent such as an ether, preferablydioxane or THF. The reaction is preferably stirred at a temperature ofapproximately 80-120° C., preferably 120° C. The reaction is preferablycarried out under an inert gas such as nitrogen or argon.

The compound of formula (C) is prepared as described above for method A.

In another embodiment, the invention relates to a process formanufacturing a compound of formula I, comprising steps a, b and c asdefined above, starting from a compound of formula (B) wherein PG¹represents R¹ (Method B-a).

The term “activating group” as used herein relates to a group that canactivate a carboxylic acid, carbonic acid or carbamic acid derivative,for coupling with an amine moiety to form an amide, urea or carbamicester moiety, respectively. Such groups are chlorides, or groupsresulting from the reaction of the acid derivative with an activatingagent. Suitable activating agents are known to the skilled person,examples of such activating reagents are carbodiimide derivatives,pentafluorophenyl ester derivatives, triazole derivatives, imidazolederivatives.

“Protecting group”:

In the methods describe above, functional groups which are present inthe starting materials and are not intended to take part in thereaction, are present in protected form if necessary, and protectinggroups that are present are cleaved, whereby said starting compounds mayalso exist in the form of salts provided that a salt-forming group ispresent and a reaction in salt form is possible. In additional processsteps, carried out as desired, functional groups of the startingcompounds which should not take part in the reaction may be present inunprotected form or may be protected for example by one or moreprotecting groups. The protecting groups are then wholly or partlyremoved according to one of the known methods.

Protecting groups, and the manner in which they are introduced andremoved are described, for example, in “Protective Groups in OrganicChemistry”, Plenum Press, London, New York 1973, and in “Methoden derorganischen Chemie”, Houben-Weyl, 4th edition, Vol. 15/1,Georg-Thieme-Verlag, Stuttgart 1974 and in Theodora W. Greene,“Protective Groups in Organic Synthesis”, John Wiley & Sons, New York1981. A characteristic of protecting groups is that they can be removedreadily, i.e. without the occurrence of undesired secondary reactions,for example by solvolysis, reduction, photolysis or alternatively underphysiological conditions.

The invention further includes any variant of the present processes, inwhich an intermediate product obtainable at any stage thereof is used asstarting material and the remaining steps are carried out, or in whichthe starting materials are formed in situ under the reaction conditions,or in which the reaction components are used in the form of their saltsor optically pure antipodes.

Compounds of the invention and intermediates can also be converted intoeach other according to methods generally known to those skilled in theart.

Intermediates and final products can be worked up and/or purifiedaccording to standard methods, e.g. using chromatographic methods,distribution methods, (re-) crystallization, and the like.

The following applies in general to all processes mentioned hereinbefore and hereinafter. All the above-mentioned process steps can becarried out under reaction conditions that are known to those skilled inthe art, including those mentioned specifically, in the absence or,customarily, in the presence of solvents or diluents, including, forexample, solvents or diluents that are inert towards the reagents usedand dissolve them, in the absence or presence of catalysts, condensationor neutralizing agents, for example ion exchangers, such as cationexchangers, e.g. in the H+ form, depending on the nature of the reactionand/or of the reactants at reduced, normal or elevated temperature, forexample in a temperature range of from about −100° C. to about 190° C.,including, for example, from approximately −80° C. to approximately 150°C., for example at from −80 to −60° C., at room temperature, at from −20to 40° C. or at reflux temperature, under atmospheric pressure or in aclosed vessel, where appropriate under pressure, and/or in an inertatmosphere, for example under an argon or nitrogen atmosphere.

At all stages of the reactions, mixtures of isomers that are formed canbe separated into the individual isomers, for example diastereoisomersor enantiomers, or into any desired mixtures of isomers, for exampleracemates or mixtures of diastereoisomers, for example analogously tothe methods described herein above.

The solvents from which those solvents that are suitable for anyparticular reaction may be selected include those mentioned specificallyor, for example, water, esters, such as lower alkyl-lower alkanoates,for example ethyl acetate, ethers, such as aliphatic ethers, for examplediethyl ether, or cyclic ethers, for example tetrahydrofuran or dioxane,liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, suchas methanol, ethanol or 1- or 2-propanol, nitriles, such asacetonitrile, halogenated hydrocarbons, such as methylene chloride orchloroform, acid amides, such as dimethylformamide or dimethylacetamide, bases, such as heterocyclic nitrogen bases, for examplepyridine or N-methylpyrrolidin-2-one, carboxylic acid anhydrides, suchas lower alkanoic acid anhydrides, for example acetic anhydride, cyclic,linear or branched hydrocarbons, such as cyclohexane, hexane orisopentane, methycyclohexane, or mixtures of those solvents, for exampleaqueous solutions, unless otherwise indicated in the description of theprocesses. Such solvent mixtures may also be used in working up, forexample by chromatography or partitioning. The compounds, includingtheir salts, may also be obtained in the form of hydrates, or theircrystals may, for example, include the solvent used for crystallization.Different crystalline forms may be present.

The invention relates also to those forms of the process in which acompound obtainable as an intermediate at any stage of the process isused as starting material and the remaining process steps are carriedout, or in which a starting material is formed under the reactionconditions or is used in the form of a derivative, for example in aprotected form or in the form of a salt, or a compound obtainable by theprocess according to the invention is produced under the processconditions and processed further in situ.

All starting materials, building blocks, reagents, acids, bases,dehydrating agents, solvents and catalysts utilized to synthesize thecompounds of the present invention are either commercially available orcan be produced by organic synthesis methods known to one of ordinaryskill in the art (Houben-Weyl 4^(th) Ed. 1952, Methods of OrganicSynthesis, Thieme, Volume 21).

Members of the phosphoinositide-3 kinase (PI3K) family are involved incell growth, differentiation, survival, cytoskeletal remodeling and thetrafficking of intracellular organelles in many different types of cells(Okkenhaug and Wymann, Nature Rev. Immunol. 3:317 (2003).

To date, eight mammalian PI3Ks have been identified, divided into threemain classes (I, II and III) on the basis of their genetic sequence,structure, adapter molecules, expression, mode of activation, andpreferred substrate.

PI3Kδ is a lipid kinase belonging to the class I PI3K family (PI3K α, β,γ and δ) that generates second messenger signals downstream of tyrosinekinase-linked receptors.

PI3Kδ is a heterodimer composed of an adaptor protein and a p110δcatalytic subunit which converts phosphatidylinositol-4,5-bis-phosphate(PtdInsP2) to phosphatidylinositol-3,4,5-tri-phosphate (PtdInsP3).Effector proteins interact with PtdInsP3 and trigger specific signalingpathways involved in cell activation, differentiation, migration, andcell survival.

Expression of the p110δ and p110γ catalytic subunits is preferential toleukocytes.

Expression is also observed in smooth muscle cells, myocytes andendothelial cells. In contrast, p110α and p110β are expressed by allcell types (Marone et al. Biochimica et Biophysica Acta 1784:159(2008)).

PI3Kδ is associated with B cell development and function (Okkenhaug etal. Science 297:1031 (2002)).

B cells play also a critical role in the pathogenesis of a number ofautoimmune and allergic diseases as well as in the process of transplantrejection (Martin and Chan, Annu. Rev. Immunol. 24:467 (2006)).

Chemotaxis is involved in many autoimmune or inflammatory diseases, inangiogenesis, invasion/metastasis, neurodegeneration or wound healing(Gerard et al. Nat. Immunol. 2:108 (2001)). Temporarily distinct eventsin leukocyte migration in response to chemokines are fully dependent onPI3Kδ and PI3Kγ (Liu et al. Blood 110:1191 (2007)).

PI3Kα and PI3Kβ play an essential role in maintaining homeostasis andpharmacological inhibition of these molecular targets has beenassociated with cancer therapy (Maira et al. Expert Opin. Ther. Targets12:223 (2008)).

PI3Kα is involved in insulin signaling and cellular growth pathways(Foukas et al. Nature 441:366 (2006)). PI3Kδ isoform-selectiveinhibition is expected to avoid potential side effects such ashyperglycemia, and metabolic or growth disregulation.

The invention relates in a third aspect to the use of compounds of thepresent invention as pharmaceuticals. Particularly, the compounds offormula I have valuable pharmacological properties, as describedhereinbefore and hereinafter. The invention thus provides:

-   -   a compound of the formula (I) as defined herein, as        pharmaceutical/for use as pharmaceutical;    -   a compound of the formula (I) as defined herein, as        medicament/for use as medicament;    -   a compound of the formula (I) as defined herein, for use in        therapy;    -   a compound of the formula (I) as defined herein, for the        prevention and/or treatment of conditions, diseases or disorders        which are mediated by the activity of the PI3K enzymes,        preferably by the activity of the PI3Kδ isoform;    -   the use of a compound of formula (I) as defined herein, for the        manufacture of a medicament for the prevention and/or treatment        of conditions, diseases or disorders which are mediated by the        activity of the PI3K enzymes, preferably by the activity of the        PI3Kδ isoform;    -   the use of a compound of formula (I) as defined herein, for the        prevention and/or treatment of conditions, diseases or disorders        which are mediated by the activity of the PI3K enzymes,        preferably by the activity of the PI3Kδ isoform;    -   the use of a compound of formula I as defined herein for the        inhibition of the PI3K, enzymes, preferably of the PI3Kδ        isoform;    -   the use of a compound of formula (I) as defined herein, for the        treatment of a disorder or disease selected from autoimmune        disorders, inflammatory diseases, allergic diseases, airway        diseases, such as asthma and COPD, transplant rejection, cancers        eg of hematopoietic origin or solid tumors.    -   the use of a compound of formula (I) as defined herein, for the        treatment of a disorder or disease selected from antibody        production, antigen presentation, cytokine production or        lymphoid organogenesis are abnormal or are undesirable including        rheumatoid arthritis, pemphigus vulgaris, idiopathic        thrombocytopenia purpura, systemic lupus erythematosus, multiple        sclerosis, myasthenia gravis, Sjögren's syndrome, autoimmune        hemolytic anemia, ANCA-associated vasculitides,        cryoglobulinemia, thrombotic thrombocytopenic purpura, chronic        autoimmune urticaria, allergy (atopic dermatitis, contact        dermatitis, allergic rhinitis), goodpasture's syndrome, AMR        (antibody-mediated transplant rejection), B cell-mediated        hyperacute, acute and chronic transplant rejection and cancers        of haematopoietic origin including but not limited to multiple        myeloma; acute myelogenous leukemia; chronic myelogenous        leukemia; lymphocytic leukemia; myeloid leukemia; non-Hodgkin        lymphoma; lymphomas; polycythemia vera; essential        thrombocythemia; myelofibrosis with myeloid metaplasia; and        Walden stroem disease.    -   the use of a compound of formula (I) as defined herein, for the        treatment of a disorder or disease selected from rheumatoid        arthritis (RA), pemphigus vulgaris (PV), idiopathic        thrombocytopenia purpura (ITP), thrombotic thrombocytopenic        purpura (TTP), autoimmune hemolytic anemia (AIHA), acquired        hemophilia type A (AHA), systemic lupus erythematosus (SLE),        multiple sclerosis (MS), myasthenia gravis (MG), Sjögren's        syndrome (SS), ANCA-associated vasculitides, cryoglobulinemia,        chronic autoimmune urticaria (CAU), allergy (atopic dermatitis,        contact dermatitis, allergic rhinitis), goodpasture's syndrome,        transplant rejection and cancers of haematopoietic origin.    -   a method of modulating the activity of the PI3K enzymes,        preferably the PI3Kδ isoform, in a subject, comprising the step        of administering to a subject a therapeutically effective amount        of a compound of formula I as defined herein;    -   a method for the treatment of a disorder or disease mediated by        the PI3K enzymes, preferably by the PI3Kδ isoform. comprising        the step of administering to a subject a therapeutically        effective amount of a compound of formula (I) as defined herein;    -   a method for inhibition of the PI3K enzymes, preferably the        PI3Kδ isoform, in a cell, comprising contacting said cell with        an effective amount of a compound of formula I as defined        herein.

As used herein, the term “subject” refers to an animal. Typically theanimal is a mammal. A subject also refers to for example, primates(e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats,mice, fish, birds and the like. In certain embodiments, the subject is aprimate. In yet other embodiments, the subject is a human.

As used herein, the term “inhibit”, “inhibition” or “inhibiting” refersto the reduction or suppression of a given condition, symptom, ordisorder, or disease, or a significant decrease in the baseline activityof a biological activity or process.

As used herein, the term “treat”, “treating” or “treatment” of anydisease or disorder refers in one embodiment, to ameliorating thedisease or disorder (i.e., slowing or arresting or reducing thedevelopment of the disease or at least one of the clinical symptomsthereof). In another embodiment “treat”, “treating” or “treatment”refers to alleviating or ameliorating at least one physical parameterincluding those which may not be discernible by the patient. In yetanother embodiment, “treat”, “treating” or “treatment” refers tomodulating the disease or disorder, either physically, (e.g.,stabilization of a discernible symptom), physiologically, (e.g.,stabilization of a physical parameter), or both. In yet anotherembodiment, “treat”, “treating” or “treatment” refers to preventing ordelaying the onset or development or progression of the disease ordisorder.

As used herein, a subject is “in need of” a treatment if such subjectwould benefit biologically, medically or in quality of life from suchtreatment.

The term “administration” or “administering” of the subject compoundmeans providing a compound of the invention and prodrugs thereof to asubject in need of treatment.

Administration “in combination with” one or more further therapeuticagents includes simultaneous (concurrent) and consecutive administrationin any order, and in any route of administration.

The invention relates to the use of new tetrahydro-pyrido-pyrimidinederivates for the prevention and/or treatment of conditions, diseases ordisorders which are mediated by the activity of the PI3K enzymes.

Suitably, the invention relates to the treatment, either alone or incombination, with one or more other pharmacologically active compounds,of PI3K-related diseases including but not limited to autoimmunedisorders, inflammatory diseases, allergic diseases, airway diseases,such as asthma and COPD, transplant rejection, cancers eg ofhematopoietic origin or solid tumors.

The invention also relates to the treatment, either alone or incombination, with one or more other pharmacologically active compounds,includes methods of treating conditions, diseases or disorders in whichone or more of the functions of B cells such as antibody production,antigen presentation, cytokine production or lymphoid organogenesis areabnormal or are undesirable including rheumatoid arthritis, pemphigusvulgaris, idiopathic thrombocytopenia purpura, systemic lupuserythematosus, multiple sclerosis, myasthenia gravis, Sjögren'ssyndrome, autoimmune hemolytic anemia, ANCA-associated vasculitides,cryoglobulinemia, thrombotic thrombocytopenic purpura, chronicautoimmune urticaria, allergy (atopic dermatitis, contact dermatitis,allergic rhinitis), goodpasture's syndrome, AMR (antibody-mediatedtransplant rejection), B cell-mediated hyperacute, acute and chronictransplant rejection and cancers of haematopoietic origin including butnot limited to multiple myeloma; acute myelogenous leukemia; chronicmyelogenous leukemia; lymphocytic leukemia; myeloid leukemia;non-Hodgkin lymphoma; lymphomas; polycythemia vera; essentialthrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenstroem disease.

The invention includes methods of treating conditions, diseases ordisorders in which one or more of the functions of neutrophils, such assuperoxide release, stimulated exocytosis, or chemoatractic migrationare abnormal or are undesirable including rheumatoid arthritis, sepsis,pulmonary or respiratory disorders such as asthma, inflammatorydermatoses such as psoriasis and others.

The invention includes methods of treating conditions, diseases ordisorders in which one or more of the functions of basophil and mastcells such as chemoatractic migration or allergen-IgE-mediateddegranulation are abnormal or are undesirable including allergicdiseases (atopic dermatitis, contact dermatitis, allergic rhinitis) aswell as other disorders such as COPD, asthma or emphysema.

The invention includes methods of treating conditions, diseases ordisorders in which one or more of the functions of T cells such ascytokine production or cell-mediated cytotoxicity abnormal or areundesirable including rheumatoid arthritis, multiple sclerosis, acute orchronic rejection of cell tissue or organ grafts or cancers ofhaematopoietic origin.

Further, the invention includes methods of treating neurodegenerativediseases, cardiovascular diseases and platelet aggregation.

Further, the invention includes methods of treating skin diseases suchas porphyria cutanea tarda, polymorphous light eruption,dermatomyositis, solar urticaria, oral lichen planus, panniculitis,scleroderma, urticarial vasculitis.

Further, the invention includes methods of treating chronic inflammatorydiseases such as sarcoidosis, granuloma annulare.

In other embodiments, the condition or disorder (e.g. PI3K-mediated) isselected from the group consisting of: polycythemia vera, essentialthrombocythemia, myelofibrosis with myeloid metaplasia, asthma, COPD,ARDS, Loffler's syndrome, eosinophilic pneumonia, parasitic (inparticular metazoan) infestation (including tropical eosinophilia),bronchopulmonary aspergillosis, polyarteritis nodosa (includingChurg-Strauss syndrome), eosinophilic granuloma, eosinophil-relateddisorders affecting the airways occasioned by drug-reaction, psoriasis,contact dermatitis, atopic dermatitis, alopecia areata, erythemamultiforme, dermatitis herpetiformis, scleroderma, vitiligo,hypersensitivity angiitis, urticaria, bullous pemphigoid, lupuserythematosus, pemphigus, epidermolysis bullosa acquisita, autoimmunehaematogical disorders (e.g. haemolytic anaemia, aplastic anaemia, purered cell anaemia and idiopathic thrombocytopenia), systemic lupuserythematosus, polychondritis, scleroderma, Wegener granulomatosis,dermatomyositis, chronic active hepatitis, myasthenia gravis,Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory boweldisease (e.g. ulcerative colitis and Crohn's disease), endocrineopthalmopathy, Grave's disease, sarcoidosis, alveolitis, chronichypersensitivity pneumonitis, multiple sclerosis, primary biliarycirrhosis, uveitis (anterior and posterior), interstitial lung fibrosis,psoriatic arthritis, glomerulonephritis, cardiovascular diseases,atherosclerosis, hypertension, deep venous thrombosis, stroke,myocardial infarction, unstable angina, thromboembolism, pulmonaryembolism, thrombolytic diseases, acute arterial ischemia, peripheralthrombotic occlusions, and coronary artery disease, reperfusioninjuries, retinopathy, such as diabetic retinopathy or hyperbaricoxygen-induced retinopathy, and conditions characterized by elevatedintraocular pressure or secretion of ocular aqueous humor, such asglaucoma.

In another embodiment, the compounds of the present invention are usefulin the treatment, prevention, or amelioration of autoimmune disease andof inflammatory conditions, in particular inflammatory conditions withan aetiology including an autoimmune component such as arthritis (forexample rheumatoid arthritis, arthritis chronica progrediente andarthritis deformans) and rheumatic diseases, including inflammatoryconditions and rheumatic diseases involving bone loss, inflammatorypain, spondyloarhropathies including ankolsing spondylitis, Reitersyndrome, reactive arthritis, psoriatic arthritis, and enterophathicsarthritis, hypersensitivity (including both airways hypersensitivity anddermal hypersensitivity) and allergies. Specific auto-immune diseasesfor which antibodies of the invention may be employed include autoimmunehaematological disorders (including e.g. hemolytic anaemia, aplasticanaemia, pure red cell anaemia and idiopa-thic thrombocytopenia),acquired hemophilia A, cold agglutinin disease, cryoglobulinemia,thrombotic thrombocytopenic purpura, Sjögren's syndrome, systemic lupuserythematosus, inflammatory muscle disorders, polychondritis,sclerodoma, anti-neutrophil cytoplasmic antibody-associated vasculitis,IgM mediated neuropathy, opsoclonus myoclonus syndrome, Wegenergranulomatosis, dermatomyositis, chronic active hepatitis, myastheniagravis, psoriasis, Steven-Johnson syndrome, pemphigus vulgaris,pemphigus foliacius, idio-pathic sprue, autoimmune inflammatory boweldisease (including e.g. ulcerative colitis, Crohn's disease andIrritable Bowel Syndrome), endocrine ophthalmopathy, Graves' disease,sarcoidosis, multiple sclerosis, neuromyelitis optica, primary biliarycirrhosis, juvenile diabetes (diabetes mellitus type I), uveitis(anterior, intermediate and posterior as well as panuveitis),keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitiallung fibrosis, psoriatic arthritis and glomerulonephritis (with andwithout nephrotic syndrome, e.g. including idiopathic nephro-ticsyndrome or minimal change nephropathy), tumors, inflammatory disease ofskin and cornea, myositis, loosening of bone implants, metabolicdisorders, such as atherosclerosis, diabetes, and dislipidemia.

In another embodiment, the compounds of the present invention are usefulin the treatment of conditions or disorders selected from the groupconsisting of, primary cutaneous B-cell lymphoma, immunobullous disease,pemphigus vulgaris, pemphigus foliaceus, paraneoplastic pemphigus,bullous pemphigoid, mucous membrane pemphigoid, epidermolysis bullosaacquisita, chronic graft versus host disease, dermatomyositis, systemiclupus erythematosus, vasculitis, small vessel vasculitis,hypocomplementemic urticarial vasculitis, antineutrophil cytoplasmicantibody-vasculitis, cryoglobulinemia, Schnitzler syndrome,Waldenstrom's macroglobulinemia, angioedema, vitiligo, systemic lupuserythematosus, idiopathic thrombocytopenic purpura, multiple sclerosis,cold agglutinin disease, autoimmune hemolytic anemia, antineutrophilcytoplasmic antibody-associated vasculitis, graft versus host disease,cryoglobulinemia and thrombotic thrombocytopenic.

In a further embodiment, the invention relates to a process or a methodfor the treatment of one of the disorders or diseases mentionedhereinabove, especially a disease which responds to the inhibition ofthe PI3K enzymes. The compounds of formula I, or a pharmaceuticallyacceptable salt thereof, can be administered as such or in the form ofpharmaceutical compositions, prophylactically or therapeutically,preferably in an amount effective against the said diseases, to awarm-blooded animal, for example a human, requiring such treatment, thecompounds especially being used in the form of pharmaceuticalcompositions.

In a further embodiment, the invention relates to the use of a compoundof formula I, or a pharmaceutically acceptable salt thereof, as such orin the form of a pharmaceutical composition with at least onepharmaceutically acceptable carrier, for the therapeutic and alsoprophylactic management of one or more of the diseases mentionedhereinabove, mediated by the PI3K enzymes.

In a further embodiment, the invention relates to the use of a compoundof formula I, or a pharmaceutically acceptable salt thereof, especiallya compound of formula I which is said to be preferred, or apharmaceutically acceptable salt thereof, for the preparation of apharmaceutical composition for the therapeutic and also prophylacticmanagement of one or more of the diseases mentioned hereinabove,especially a disorder or disease selected from autoimmune disorders,inflammatory diseases, allergic diseases, airway diseases, such asasthma and COPD, transplant rejection, cancers eg of hematopoieticorigin or solid tumors.

The invention relates in a fourth aspect to pharmaceutical compositionscomprising a compound of the present invention. The invention thusprovides

-   -   a pharmaceutical composition comprising (i.e. containing or        consisting of) a compound as defined herein and one or more        carriers/excipients;    -   a pharmaceutical composition comprising a therapeutically        effective amount of a compound of formula I as defined herein,        and one or more pharmaceutically acceptable carriers/excipients.

As used herein, the term “pharmaceutically acceptable carrier” includesany and all solvents, dispersion media, coatings, surfactants,antioxidants, preservatives (e.g., antibacterial agents, antifungalagents), isotonic agents, absorption delaying agents, salts,preservatives, drugs, drug stabilizers, binders, excipients,disintegration agents, lubricants, sweetening agents, flavoring agents,dyes, and the like and combinations thereof, as would be known to thoseskilled in the art (see, for example, Remington's PharmaceuticalSciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329). Exceptinsofar as any conventional carrier is incompatible with the activeingredient, its use in the therapeutic or pharmaceutical compositions iscontemplated.

The present invention provides a pharmaceutical composition comprising acompound of the present invention and a pharmaceutically acceptablecarrier. The pharmaceutical composition can be formulated for particularroutes of administration such as oral administration, parenteraladministration, and rectal administration, etc. In addition, thepharmaceutical compositions of the present invention can be made up in asolid form (including without limitation capsules, tablets, pills,granules, powders or suppositories), or in a liquid form (includingwithout limitation solutions, suspensions or emulsions). Thepharmaceutical compositions can be subjected to conventionalpharmaceutical operations such as sterilization and/or can containconventional inert diluents, lubricating agents, or buffering agents, aswell as adjuvants, such as preservatives, stabilizers, wetting agents,emulsifiers and buffers, etc.

Typically, the pharmaceutical compositions are tablets or gelatincapsules comprising the active ingredient together with

-   -   a) diluents, e.g., lactose, dextrose, sucrose, mannitol,        sorbitol, cellulose and/or glycine;    -   b) lubricants, e.g., silica, talcum, stearic acid, its magnesium        or calcium salt and/or polyethyleneglycol; for tablets also    -   c) binders, e.g., magnesium aluminum silicate, starch paste,        gelatin, tragacanth, methylcellulose, sodium        carboxymethylcellulose and/or polyvinylpyrrolidone; if desired    -   d) disintegrants, e.g., starches, agar, alginic acid or its        sodium salt, or effervescent mixtures; and/or    -   e) absorbents, colorants, flavors and sweeteners.

Tablets may be either film coated or enteric coated according to methodsknown in the art. Suitable compositions for oral administration includean effective amount of a compound of the invention in the form oftablets, lozenges, aqueous or oily suspensions, dispersible powders orgranules, emulsion, hard or soft capsules, or syrups or elixirs.Compositions intended for oral use are prepared according to any methodknown in the art for the manufacture of pharmaceutical compositions andsuch compositions can contain one or more agents selected from the groupconsisting of sweetening agents, flavoring agents, coloring agents andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets may contain the active ingredient inadmixture with nontoxic pharmaceutically acceptable excipients which aresuitable for the manufacture of tablets. These excipients are, forexample, inert diluents, such as calcium carbonate, sodium carbonate,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example, starch, gelatin or acacia; and lubricatingagents, for example magnesium stearate, stearic acid or talc. Thetablets are uncoated or coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearate canbe employed. Formulations for oral use can be presented as hard gelatincapsules wherein the active ingredient is mixed with an inert soliddiluent, for example, calcium carbonate, calcium phosphate or kaolin, oras soft gelatin capsules wherein the active ingredient is mixed withwater or an oil medium, for example, peanut oil, liquid paraffin orolive oil.

Certain injectable compositions are aqueous isotonic solutions orsuspensions, and suppositories are advantageously prepared from fattyemulsions or suspensions. Said compositions may be sterilized and/orcontain adjuvants, such as preserving, stabilizing, wetting oremulsifying agents, solution promoters, salts for regulating the osmoticpressure and/or buffers. In addition, they may also contain othertherapeutically valuable substances. Said compositions are preparedaccording to conventional mixing, granulating or coating methods,respectively, and contain about 0.1-75%, or contain about 1-50%, of theactive ingredient.

Suitable compositions for transdermal application include an effectiveamount of a compound of the invention with a suitable carrier. Carrierssuitable for transdermal delivery include absorbable pharmacologicallyacceptable solvents to assist passage through the skin of the host. Forexample, transdermal devices are in the form of a bandage comprising abacking member, a reservoir containing the compound optionally withcarriers, optionally a rate controlling barrier to deliver the compoundof the skin of the host at a controlled and predetermined rate over aprolonged period of time, and means to secure the device to the skin.

Suitable compositions for topical application, e.g., to the skin andeyes, include aqueous solutions, suspensions, ointments, creams, gels orsprayable formulations, e.g., for delivery by aerosol or the like. Suchtopical delivery systems will in particular be appropriate for dermalapplication, e.g., for the treatment of skin cancer, e.g., forprophylactic use in sun creams, lotions, sprays and the like. They arethus particularly suited for use in topical, including cosmetic,formulations well-known in the art. Such may contain solubilizers,stabilizers, tonicity enhancing agents, buffers and preservatives.

As used herein a topical application may also pertain to an inhalationor to an intranasal application. They may be conveniently delivered inthe form of a dry powder, either alone, as a mixture, for example a dryblend with lactose, or a mixed component particle, for example withphospholipids, from a dry powder inhaler or an aerosol spraypresentation from a pressurised container, pump, spray, atomizer ornebuliser, with or without the use of a suitable propellant.

The present invention further provides anhydrous pharmaceuticalcompositions and dosage forms comprising the compounds of the presentinvention as active ingredients, since water may facilitate thedegradation of certain compounds.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. An anhydrous pharmaceuticalcomposition may be prepared and stored such that its anhydrous nature ismaintained. Accordingly, anhydrous compositions are packaged usingmaterials known to prevent exposure to water such that they can beincluded in suitable formulary kits. Examples of suitable packaginginclude, but are not limited to, hermetically sealed foils, plastics,unit dose containers, e. g., vials, blister packs, and strip packs.

The invention further provides pharmaceutical compositions and dosageforms that comprise one or more agents that reduce the rate by which thecompound of the present invention as an active ingredient willdecompose. Such agents, which are referred to herein as “stabilizers,”include, but are not limited to, antioxidants such as ascorbic acid, pHbuffers, or salt buffers, etc.

Examples of physiologically acceptable carriers include buffers such asphosphate, citrate, and other organic acids; antioxidants includingascorbic acid; low molecular weight (less than about 10 residues)polypeptide; proteins, such as serum albumin, gelatin, orimmunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone;amino acids such as glycine, glutamine, asparagine, arginine or lysine;monosaccharides, disaccharides, and other carbohydrates includingglucose, mannose, or dextrins; chelating agents such as EDTA; sugaralcohols such as mannitol or sorbitol; salt-forming counterions such assodium; and/or nonionic surfactants such as TWEEN®, polyethylene glycol(PEG), and PLURONICS®.

Suitable excipients/carriers may be any solid, liquid, semi-solid or, inthe case of an aerosol composition, gaseous excipient that is generallyavailable to one of skill in the art.

Solid pharmaceutical excipients include starch, cellulose, talc,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, magnesium stearate, sodium stearate, glycerol monostearate, sodiumchloride, dried skim milk and the like.

Liquid and semisolid excipients may be selected from glycerol, propyleneglycol, water, ethanol and various oils, including those of petroleum,animal, vegetable or synthetic origin, e.g., peanut oil, soybean oil,mineral oil, sesame oil, etc. Preferred liquid carriers, particularlyfor injectable solutions, include water, saline, aqueous dextrose, andglycols.

Compressed gases may be used to disperse a compound of the formula (I)in aerosol form. Inert gases suitable for this purpose are nitrogen,carbon dioxide, etc. Other suitable pharmaceutical excipients and theirformulations are described in Remington's Pharmaceutical Sciences,edited by E. W. Martin (Mack Publishing Company, 18th ed., 1990).

The dosage of the active ingredient depends upon the disease to betreated and upon the species, its age, weight, and individual condition,the individual pharmacokinetic data, and the mode of administration. Theamount of the compound in a formulation can vary within the full rangeemployed by those skilled in the art. Typically, the formulation willcontain, on a weight percent (wt %) basis, from about 0.01-99.99 wt % ofa compound of formula (I) based on the total formulation, with thebalance being one or more suitable pharmaceutical excipients.

Pharmaceutical compositions comprising a compound of formula (I) asdefined herein in association with at least one pharmaceuticalacceptable carrier (such as excipient a and/or diluent) may bemanufactured in conventional manner, e.g. by means of conventionalmixing, granulating, coating, dissolving or lyophilising processes.

In a further embodiment, the invention relates to a pharmaceuticalcomposition for administration to a warm-blooded animal, especiallyhumans or commercially useful mammals suffering from a disease whichresponds to an inhibition of the PI3K enzymes, comprising an effectivequantity of a compound of formula I for the inhibition of the PI3Kenzymes, or a pharmaceutically acceptable salt thereof, together with atleast one pharmaceutically acceptable carrier.

In a further embodiment, the invention relates to a pharmaceuticalcomposition for the prophylactic or especially therapeutic management ofa disorder or disease selected from autoimmune disorders, inflammatorydiseases, allergic diseases, airway diseases, such as asthma and COPD,transplant rejection, cancers eg of hematopoietic origin or solidtumors; of a warm-blooded animal, especially a human or a commerciallyuseful mammal requiring such treatment.

The invention relates in a fifth aspect to combinations comprising acompound of formula I and one or more additional active ingredients. Theinvention thus provides

-   -   a combination in particular a pharmaceutical combination,        comprising a therapeutically effective amount of a compound of        formula I and one or more therapeutically active agents, e.g. an        immunosuppressant, immunomodulatory, anti-inflammatory or        chemotherapeutic agent, e.g. as indicated below;    -   a combined pharmaceutical composition, adapted for simultaneous        or sequential administration, comprising a therapeutically        effective amount of a compound of formula (I) as defined herein;        therapeutically effective amount(s) of one or more combination        partners e.g. an immunosuppressant, immunomodulatory,        anti-inflammatory or chemotherapeutic agent, e.g. as indicated        below; one or more pharmaceutically acceptable excepients;    -   a combined pharmaceutical composition as defined herein (i) as        pharmaceutical, (ii) for use in the treatment of a disease        mediated by the PI3K enzymes, (iii) in a method of treatment of        a disease mediated by the PI3K enzymes.

By “combination”, there is meant either a fixed combination in onedosage unit form, or a kit of parts for the combined administrationwhere a compound of the formula (I) and a combination partner may beadministered independently at the same time or separately within timeintervals that especially allow that the combination partners show acooperative, e.g. synergistic effect.

The term “a therapeutically effective amount” of a compound of thepresent invention refers to an amount of the compound of the presentinvention that will elicit the biological or medical response of asubject, for example, reduction or inhibition of an enzyme or a proteinactivity, or ameliorate symptoms, alleviate conditions, slow or delaydisease progression, or prevent a disease, etc. In one non-limitingembodiment, the term “a therapeutically effective amount” refers to theamount of the compound of the present invention that, when administeredto a subject, is effective to (1) at least partially alleviating,inhibiting, preventing and/or ameliorating a condition, or a disorder ora disease (i) mediated by the dysregulation of PI3K delta, or (ii)associated with the dysregulation of PI3K delta, or (iii) characterizedby the dysregulation of the PI3K delta; or (2) reducing or inhibitingthe activity of the PI3K delta. In another non-limiting embodiment, theterm “a therapeutically effective amount” refers to the amount of thecompound of the present invention that, when administered to a cell, ora tissue, or a non-cellular biological material, or a medium, iseffective to at least partially reducing or inhibiting PI3K delta.

The compounds of formula I may be administered as the sole activeingredient or in conjunction with, e.g. as an adjuvant to, other drugse.g. immunosuppressive or immunomodulating agents or otheranti-inflammatory agents, e.g. for the treatment or prevention of allo-or xenograft acute or chronic rejection or inflammatory or autoimmunedisorders, or a chemotherapeutic agent, e.g a malignant cellanti-proliferative agent. For example, the compounds of formula I may beused in combination with a calcineurin inhibitor, e.g. cyclosporin A orFK 506; a mTOR inhibitor, e.g. rapamycin,40-O-(2-hydroxyethyl)-rapamycin, CC1779, ABT578, AP23573, TAFA-93,biolimus-7 or biolimus-9; an ascomycin having immuno-suppressiveproperties, e.g. ABT-281, ASM981, etc.; corticosteroids;cyclophosphamide; azathioprene; methotrexate; leflunomide; mizoribine;mycophenolic acid or salt; mycophenolate mofetil; 15-deoxyspergualine oran immunosuppressive homologue, analogue or derivative thereof; a PKCinhibitor, e.g. as disclosed in WO 02/38561 or WO 03/82859, e.g. thecompound of Example 56 or 70; a JAK3 kinase inhibitor, e.g.N-benzyl-3,4-dihydroxy-benzylidene-cyanoacetamideα-cyano-(3,4-dihydroxy)-]N-benzylcinnamamide (Tyrphostin AG 490),prodigiosin 25-C (PNU156804),[4-(4′-hydroxyphenyl)-amino-6,7-dimethoxyquinazoline] (WHI-P131),[4-(3′-bromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline](WHI-P154),[4-(3′,5′-dibromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline]WHI-P97, KRX-211,3-{(3R,4R)-4-methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile,in free form or in a pharmaceutically acceptable salt form, e.g.mono-citrate (also called CP-690,550), or a compound as disclosed in WO04/052359 or WO 05/066156; immunosuppressive monoclonal antibodies,e.g., monoclonal antibodies to leukocyte receptors, e.g., MHC, CD2, CD3,CD4, CD7, CD8, CD25, CD28, CD40, CD45, CD52, CD58, CD80, CD86 or theirligands; other immunomodulatory compounds, e.g. a recombinant bindingmolecule having at least a portion of the extracellular domain of CTLA4or a mutant thereof, e.g. an at least extracellular portion of CTLA4 ora mutant thereof joined to a non-CTLA4 protein sequence, e.g. CTLA4lg(for ex. designated ATCC 68629) or a mutant thereof, e.g. LEA29Y;adhesion molecule inhibitors, e.g. LFA-1 antagonists, ICAM-1 or -3antagonists, VCAM-4 antagonists or VLA-4 antagonists; or antihistamines;or antitussives, or a bronchodilatory agent; or an angiotensin receptorblockers; or an anti-infectious agent.

Where the compounds of formula I are administered in conjunction withother immunosuppressive/immunomodulatory, anti-inflammatory,chemotherapeutic or anti-infectious therapy, dosages of theco-administered immunosuppressant, immunomodulatory, anti-inflammatory,chemotherapeutic or anti-infectious compound will of course varydepending on the type of co-drug employed, e.g. whether it is a steroidor a calcineurin inhibitor, on the specific drug employed, on thecondition being treated and so forth.

A compound of the formula (I) may also be used to advantage incombination with each other or in combination with other therapeuticagents, especially other antiproliferative agents. Suchantiproliferative agents include, but are not limited to, aromataseinhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase IIinhibitors; microtubule active agents; alkylating agents; histonedeacetylase inhibitors; compounds, which induce cell differentiationprocesses; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors;antineoplastic antimetabolites; platin compounds; compoundstargeting/decreasing a protein or lipid kinase activity and furtheranti-angiogenic compounds; compounds which target, decrease or inhibitthe activity of a protein or lipid phosphatase; gonadorelin agonists;anti-androgens; methionine aminopeptidase inhibitors; bisphosphonates;biological response modifiers; antiproliferative antibodies; heparanaseinhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors;proteasome inhibitors; agents used in the treatment of hematologicmalignancies; compounds which target, decrease or inhibit the activityof Flt-3; Hsp90 inhibitors; temozolomide (TEMODAL®); and leucovorin.

The term “aromatase inhibitor”, as used herein, relates to a compoundwhich inhibits the estrogen production, i.e., the conversion of thesubstrates androstenedione and testosterone to estrone and estradiol,respectively. The term includes, but is not limited to, steroids,especially atamestane, exemestane and formestane; and, in particular,non-steroids, especially aminoglutethimide, roglethimide,pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole,fadrozole, anastrozole and letrozole. Exemestane can be administered,e.g., in the form as it is marketed, e.g., under the trademark AROMASIN.Formestane can be administered, e.g., in the form as it is marketed,e.g., under the trademark LENTARON. Fadrozole can be administered, e.g.,in the form as it is marketed, e.g., under the trademark AFEMA.Anastrozole can be administered, e.g., in the form as it is marketed,e.g., under the trademark ARIMIDEX. Letrozole can be administered, e.g.,in the form as it is marketed, e.g., under the trademark FEMARA orFEMAR. Aminoglutethimide can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ORIMETEN. A combination of theinvention comprising a chemotherapeutic agent which is an aromataseinhibitor is particularly useful for the treatment of hormone receptorpositive tumors, e.g., breast tumors.

The term “anti-estrogen”, as used herein, relates to a compound whichantagonizes the effect of estrogens at the estrogen receptor level. Theterm includes, but is not limited to, tamoxifen, fulvestrant, raloxifeneand raloxifene hydrochloride. Tamoxifen can be administered, e.g., inthe form as it is marketed, e.g., under the trademark NOLVADEX.Raloxifene hydrochloride can be administered, e.g., in the form as it ismarketed, e.g., under the trademark EVISTA. Fulvestrant can beformulated as disclosed in U.S. Pat. No. 4,659,516 or it can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark FASLODEX. A combination of the invention comprising achemotherapeutic agent which is an antiestrogen is particularly usefulfor the treatment of estrogen receptor positive tumors, e.g., breasttumors.

The term “anti-androgen”, as used herein, relates to any substance whichis capable of inhibiting the biological effects of androgenic hormonesand includes, but is not limited to, bicalutamide (CASODEX), which canbe formulated, e.g., as disclosed in U.S. Pat. No. 4,636,505.

The term “gonadorelin agonist”, as used herein, includes, but is notlimited to, abarelix, goserelin and goserelin acetate. Goserelin isdisclosed in U.S. Pat. No. 4,100,274 and can be administered, e.g., inthe form as it is marketed, e.g., under the trademark ZOLADEX. Abarelixcan be formulated, e.g., as disclosed in U.S. Pat. No. 5,843,901.

The term “topoisomerase I inhibitor”, as used herein, includes, but isnot limited to, topotecan, gimatecan, irinotecan, camptothecian and itsanalogues, 9-nitrocamptothecin and the macromolecular camptothecinconjugate PNU-166148 (compound A1 in WO 99/17804). Irinotecan can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark CAMPTOSAR. Topotecan can be administered, e.g., in the form asit is marketed, e.g., under the trademark HYCAMTIN.

The term “topoisomerase II inhibitor”, as used herein, includes, but isnot limited to, the anthracyclines, such as doxorubicin, includingliposomal formulation, e.g., CAELYX; daunorubicin; epirubicin;idarubicin; nemorubicin; the anthraquinones mitoxantrone andlosoxantrone; and the podophillotoxines etoposide and teniposide.Etoposide can be administered, e.g., in the form as it is marketed,e.g., under the trademark ETOPOPHOS. Teniposide can be administered,e.g., in the form as it is marketed, e.g., under the trademark VM26-BRISTOL. Doxorubicin can be administered, e.g., in the form as it ismarketed, e.g., under the trademark ADRIBLASTIN or ADRIAMYCIN.Epirubicin can be administered, e.g., in the form as it is marketed,e.g., under the trademark FARMORUBICIN. Idarubicin can be administered,e.g., in the form as it is marketed, e.g., under the trademark ZAVEDOS.Mitoxantrone can be administered, e.g., in the form as it is marketed,e.g., under the trademark NOVANTRON.

The term “microtubule active agent” relates to microtubule stabilizing,microtubule destabilizing agents and microtublin polymerizationinhibitors including, but not limited to, taxanes, e.g., paclitaxel anddocetaxel; vinca alkaloids, e.g., vinblastine, especially vinblastinesulfate; vincristine, especially vincristine sulfate and vinorelbine;discodermolides; cochicine; and epothilones and derivatives thereof,e.g., epothilone B or D or derivatives thereof. Paclitaxel may beadministered, e.g., in the form as it is marketed, e.g., TAXOL.Docetaxel can be administered, e.g., in the form as it is marketed,e.g., under the trademark TAXOTERE. Vinblastine sulfate can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark VINBLASTIN R.P. Vincristine sulfate can be administered, e.g.,in the form as it is marketed, e.g., under the trademark FARMISTIN.Discodermolide can be obtained, e.g., as disclosed in U.S. Pat. No.5,010,099. Also included are epothilone derivatives which are disclosedin WO 98/10121, U.S. Pat. No. 6,194,181, WO 98/25929, WO 98/08849, WO99/43653, WO 98/22461 and WO 00/31247. Especially preferred areepothilone A and/or B.

The term “alkylating agent”, as used herein, includes, but is notlimited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNUor Gliadel).

Cyclophosphamide can be administered, e.g., in the form as it ismarketed, e.g., under the trademark CYCLOSTIN. Ifosfamide can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark HOLOXAN.

The term “histone deacetylase inhibitors” or “HDAC inhibitors” relatesto compounds which inhibit the histone deacetylase and which possessantiproliferative activity. This includes compounds disclosed in WO02/22577, especiallyN-hydroxy-3-[4-[[(2-hydroxyethyl)[2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide,N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamideand pharmaceutically acceptable salts thereof. It further especiallyincludes suberoylanilide hydroxamic acid (SAHA).

The term “antineoplastic antimetabolite” includes, but is not limitedto, 5-fluorouracil or 5-FU; capecitabine; gemcitabine; DNA demethylatingagents, such as 5-azacytidine and decitabine; methotrexate andedatrexate; and folic acid antagonists, such as pemetrexed. Capecitabinecan be administered, e.g., in the form as it is marketed, e.g., underthe trademark XELODA. Gemcitabine can be administered, e.g., in the formas it is marketed, e.g., under the trademark GEMZAR. Also included isthe monoclonal antibody trastuzumab which can be administered, e.g., inthe form as it is marketed, e.g., under the trademark HERCEPTIN.

The term “platin compound”, as used herein, includes, but is not limitedto, carboplatin, cis-platin, cisplatinum and oxaliplatin. Carboplatincan be administered, e.g., in the form as it is marketed, e.g., underthe trademark CARBOPLAT. Oxaliplatin can be administered, e.g., in theform as it is marketed, e.g., under the trademark ELOXATIN.

The term “compounds targeting/decreasing a protein or lipid kinaseactivity; or a protein or lipid phosphatase activity; or furtheranti-angiogenic compounds”, as used herein, includes, but is not limitedto, protein tyrosine kinase and/or serine and/or threonine kinaseinhibitors or lipid kinase inhibitors, e.g.,

-   -   a) compounds targeting, decreasing or inhibiting the activity of        the platelet-derived growth factor-receptors (PDGFR), such as        compounds which target, decrease or inhibit the activity of        PDGFR, especially compounds which inhibit the PDGF receptor,        e.g., a N-phenyl-2-pyrimidine-amine derivative, e.g., imatinib,        SU101, SU6668 and GFB-111;    -   b) compounds targeting, decreasing or inhibiting the activity of        the fibroblast growth factor-receptors (FGFR);    -   c) compounds targeting, decreasing or inhibiting the activity of        the insulin-like growth factor receptor I (IGF-IR), such as        compounds which target, decrease or inhibit the activity of        IGF-IR, especially compounds which inhibit the IGF-IR receptor,        such as those compounds disclosed in WO 02/092599;    -   d) compounds targeting, decreasing or inhibiting the activity of        the Trk receptor tyrosine kinase family;    -   e) compounds targeting, decreasing or inhibiting the activity of        the Axl receptor tyrosine kinase family;    -   f) compounds targeting, decreasing or inhibiting the activity of        the c-Met receptor;    -   g) compounds targeting, decreasing or inhibiting the activity of        the Kit/SCFR receptor tyrosine kinase;    -   h) compounds targeting, decreasing or inhibiting the activity of        the C-kit receptor tyrosine kinases—(part of the PDGFR family),        such as compounds which target, decrease or inhibit the activity        of the c-Kit receptor tyrosine kinase family, especially        compounds which inhibit the c-Kit receptor, e.g., imatinib;    -   i) compounds targeting, decreasing or inhibiting the activity of        members of the c-Abl family and their gene-fusion products,        e.g., BCR-Abl kinase, such as compounds which target decrease or        inhibit the activity of c-Abl family members and their gene        fusion products, e.g., a N-phenyl-2-pyrimidine-amine derivative,        e.g., imatinib, PD180970, AG957, NSC 680410 or PD173955 from        ParkeDavis;    -   j) compounds targeting, decreasing or inhibiting the activity of        members of the protein kinase C (PKC) and Raf family of        serine/threonine kinases, members of the MEK, SRC, JAK, FAK, PDK        and Ras/MAPK family members, or PI(3) kinase family, or of the        PI(3)-kinase-related kinase family, and/or members of the        cyclin-dependent kinase family (CDK) and are especially those        staurosporine derivatives disclosed in U.S. Pat. No. 5,093,330,        e.g., midostaurin; examples of further compounds include, e.g.,        UCN-01; safingol; BAY 43-9006; Bryostatin 1; Perifosine;        Ilmofosine; RO 318220 and RO 320432; GO 6976; Isis 3521;        LY333531/LY379196; isochinoline compounds, such as those        disclosed in WO 00/09495; FTIs; PD184352; or QAN697 (a PI3K        inhibitor);    -   k) compounds targeting, decreasing or inhibiting the activity of        protein-tyrosine kinase inhibitors, such as compounds which        target, decrease or inhibit the activity of protein-tyrosine        kinase inhibitors include imatinib mesylate (GLEEVEC) or        tyrphostin. A tyrphostin is preferably a low molecular weight        (Mr<1500) compound, or a pharmaceutically acceptable salt        thereof, especially a compound selected from the        benzylidenemalonitrile class or the S-arylbenzenemalonirile or        bisubstrate quinoline class of compounds, more especially any        compound selected from the group consisting of Tyrphostin        A23/RG-50810, AG 99, Tyrphostin AG 213, Tyrphostin AG 1748,        Tyrphostin AG 490, Tyrphostin B44, Tyrphostin B44 (+)        enantiomer, Tyrphostin AG 555, AG 494, Tyrphostin AG 556, AG957        and adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic        acid adamantyl ester, NSC 680410, adaphostin; and    -   l) compounds targeting, decreasing or inhibiting the activity of        the epidermal growth factor family of receptor tyrosine kinases        (EGFR, ErbB2, ErbB3, ErbB4 as homo- or hetero-dimers), such as        compounds which target, decrease or inhibit the activity of the        epidermal growth factor receptor family are especially        compounds, proteins or antibodies which inhibit members of the        EGF receptor tyrosine kinase family, e.g., EGF receptor, ErbB2,        ErbB3 and ErbB4 or bind to EGF or EGF related ligands, and are        in particular those compounds, proteins or monoclonal antibodies        generically and specifically disclosed in WO 97/02266, e.g., the        compound of Example 39, or in EP 0 564 409; WO 99/03854; EP        0520722; EP 0 566 226; EP 0 787 722; EP 0 837 063; U.S. Pat. No.        5,747,498; WO 98/10767; WO 97/30034; WO 97/49688; WO 97/38983        and, especially, WO 96/30347, e.g., compound known as CP 358774;        WO 96/33980, e.g., compound ZD 1839; and WO 95/03283, e.g.,        compound ZM105180, e.g., trastuzumab (HERCEPTIN), cetuximab,        Iressa, Tarceva, OSI-774, CI-1033, EKB-569, GW-2016, E1.1, E2.4,        E2.5, E6.2, E6.4, E2.11, E6.3 or E7.6.3; and        7H-pyrrolo-[2,3-d]pyrimidine derivatives which are disclosed in        WO 03/013541.

Further anti-angiogenic compounds include compounds having anothermechanism for their activity, e.g., unrelated to protein or lipid kinaseinhibition, e.g., thalidomide (THALOMID) and TNP-470.

Compounds which target, decrease or inhibit the activity of a protein orlipid phosphatase are, e.g., inhibitors of phosphatase 1, phosphatase2A, PTEN or CDC25, e.g., okadaic acid or a derivative thereof.

Compounds which induce cell differentiation processes are e.g. retinoicacid, α- γ- or δ-tocopherol or α- γ- or δ-tocotrienol.

The term cyclooxygenase inhibitor, as used herein, includes, but is notlimited to, e.g., Cox-2 inhibitors, 5-alkyl substituted2-arylaminophenylacetic acid and derivatives, such as celecoxib(CELEBREX), rofecoxib (VIOXX), etoricoxib, valdecoxib or a5-alkyl-2-arylaminophenylacetic acid, e.g.,5-methyl-2-(2′-chloro-6′-fluoroanilino)phenyl acetic acid orlumiracoxib.

The term “bisphosphonates”, as used herein, includes, but is not limitedto, etridonic, clodronic, tiludronic, pamidronic, alendronic,ibandronic, risedronic and zoledronic acid. “Etridonic acid” can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark DIDRONEL. “Clodronic acid” can be administered, e.g., in theform as it is marketed, e.g., under the trademark BONEFOS. “Tiludronicacid” can be administered, e.g., in the form as it is marketed, e.g.,under the trademark SKELID. “Pamidronic acid” can be administered, e.g.,in the form as it is marketed, e.g., under the trademark AREDIA™“Alendronic acid” can be administered, e.g., in the form as it ismarketed, e.g., under the trademark FOSAMAX. “Ibandronic acid” can beadministered, e.g., in the form as it is marketed, e.g., under thetrademark BONDRANAT. “Risedronic acid” can be administered, e.g., in theform as it is marketed, e.g., under the trademark ACTONEL. “Zoledronicacid” can be administered, e.g., in the form as it is marketed, e.g.,under the trademark ZOMETA.

The term “mTOR inhibitors” relates to compounds which inhibit themammalian target of rapamycin (mTOR) and which possess antiproliferativeactivity, such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779and ABT578.

The term “heparanase inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit heparin sulphate degradation. The termincludes, but is not limited to, PI-88.

The term “biological response modifier”, as used herein, refers to alymphokine or interferons, e.g., interferon γ.

The term “inhibitor of Ras oncogenic isoforms”, e.g., H-Ras, K-Ras orN-Ras, as used herein, refers to compounds which target, decrease orinhibit the oncogenic activity of Ras, e.g., a “farnesyl transferaseinhibitor”, e.g., L-744832, DK8G557 or R115777 (Zarnestra).

The term “telomerase inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit the activity of telomerase. Compoundswhich target, decrease or inhibit the activity of telomerase areespecially compounds which inhibit the telomerase receptor, e.g.,telomestatin.

The term “methionine aminopeptidase inhibitor”, as used herein, refersto compounds which target, decrease or inhibit the activity ofmethionine aminopeptidase. Compounds which target, decrease or inhibitthe activity of methionine aminopeptidase are, e.g., bengamide or aderivative thereof.

The term “proteasome inhibitor”, as used herein, refers to compoundswhich target, decrease or inhibit the activity of the proteasome.Compounds which target, decrease or inhibit the activity of theproteasome include, e.g., PS-341 and MLN 341.

The term “matrix metalloproteinase inhibitor” or “MMP inhibitor”, asused herein, includes, but is not limited to, collagen peptidomimeticand nonpeptidomimetic inhibitors, tetracycline derivatives, e.g.,hydroxamate peptidomimetic inhibitor batimastat and its orallybioavailable analogue marimastat (BB-2516), prinomastat (AG3340),metastat (NSC 683551) BMS-279251, BAY 12-9566, TAA211, MMI270B orAAJ996.

The term “agents used in the treatment of hematologic malignancies”, asused herein, includes, but is not limited to, FMS-like tyrosine kinaseinhibitors, e.g., compounds targeting, decreasing or inhibiting theactivity of FMS-like tyrosine kinase receptors (Flt-3R); interferon,1-b-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors,e.g., compounds which target, decrease or inhibit anaplastic lymphomakinase.

Compounds which target, decrease or inhibit the activity of FMS-liketyrosine kinase receptors (Flt-3R) are especially compounds, proteins orantibodies which inhibit members of the Flt-3R receptor kinase family,e.g., PKC412, midostaurin, a staurosporine derivative, SU11248 andMLN518.

The term “HSP90 inhibitors”, as used herein, includes, but is notlimited to, compounds targeting, decreasing or inhibiting the intrinsicATPase activity of HSP90; degrading, targeting, decreasing or inhibitingthe HSP90 client proteins via the ubiquitin proteasome pathway.Compounds targeting, decreasing or inhibiting the intrinsic ATPaseactivity of HSP90 are especially compounds, proteins or antibodies whichinhibit the ATPase activity of HSP90, e.g., 17-allylamino,17-demethoxygeldanamycin (17AAG), a geldanamycin derivative, othergeldanamycin related compounds, radicicol and HDAC inhibitors.

The term “antiproliferative antibodies”, as used herein, includes, butis not limited to, trastuzumab (Herceptin™), Trastuzumab-DM1, erlotinib(Tarceva™), bevacizumab (Avastin™), rituximab (Rituxan®), PR064553(anti-CD40) and 2C4 antibody. By antibodies is meant, e.g., intactmonoclonal antibodies, polyclonal antibodies, multispecific antibodiesformed from at least two intact antibodies, and antibodies fragments solong as they exhibit the desired biological activity.

For the treatment of acute myeloid leukemia (AML), compounds of formula(I) can be used in combination with standard leukemia therapies,especially in combination with therapies used for the treatment of AML.In particular, compounds of formula (I) can be administered incombination with, e.g., farnesyl transferase inhibitors and/or otherdrugs useful for the treatment of AML, such as Daunorubicin, Adriamycin,Ara-C, VP-16, Teniposide, Mitoxantrone, Idarubicin, Carboplatinum andPKC412.

The structure of the active agents identified by code nos., generic ortrade names may be taken from the actual edition of the standardcompendium “The Merck Index” or from databases, e.g., PatentsInternational, e.g., IMS World Publications.

The above-mentioned compounds, which can be used in combination with acompound of the formula (I), can be prepared and administered asdescribed in the art, such as in the documents cited above.

A compound of the formula (I) may also be used to advantage incombination with known therapeutic processes, e.g., the administrationof hormones or especially radiation.

A compound of formula (I) may in particular be used as aradiosensitizer, especially for the treatment of tumors which exhibitpoor sensitivity to radiotherapy.

By “combination”, there is meant either a fixed combination in onedosage unit form, or a kit of parts for the combined administrationwhere a compound of the formula (I) and a combination partner may beadministered independently at the same time or separately within timeintervals that especially allow that the combination partners show acooperative, e.g., synergistic, effect or any combination thereof. Theterms “co-administration” or “combined administration” or the like asutilized herein are meant to encompass administration of the selectedcombination partner to a single subject in need thereof (e.g. apatient), and are intended to include treatment regimens in which theagents are not necessarily administered by the same route ofadministration or at the same time. The term “pharmaceuticalcombination” as used herein means a product that results from the mixingor combining of more than one active ingredient and includes both fixedand non-fixed combinations of the active ingredients. The term “fixedcombination” means that the active ingredients, e.g. a compound offormula I and a combination partner, are both administered to a patientsimultaneously in the form of a single entity or dosage. The term“non-fixed combination” means that the active ingredients, e.g. acompound of formula (I) and a combination partner, are both administeredto a patient as separate entities either simultaneously, concurrently orsequentially with no specific time limits, wherein such administrationprovides therapeutically effective levels of the two compounds in thebody of the patient. The latter also applies to cocktail therapy, e.g.the administration of three or more active ingredients.

Experimental Details:

Insofar as the production of the starting materials is not particularlydescribed, the compounds are known or may be prepared analogously tomethods known in the art or as described hereafter.

The following examples are illustrative of the invention without anylimitation.

Abbreviations:

-   AcOH acetic acid-   aq aqueous-   Ar aryl-   BOC tert-butyl-carbonate-   BOP benzotriazol-1-yloxytris(dimethylamino)phosphonium    hexafluorophosphate-   br. s. broad singlet-   CDCl₃ chloroform-d-   CDI 1,1′-carbonyldiimidazole-   CH₂Cl₂ dichloromethane-   CH₃CN acetonitrile-   Cs₂CO₃ cesium carbonate-   d doublet-   dd doublet of doublets-   DIPEA N-ethyldiisopropylamine-   DME 1,4-dimethoxyethane-   DMF N,N-dimethylformamide-   DBU 1,8-diaza-7-bicyclo[5.4.0]undecene-   DMSO dimethylsulfoxide-   dt doublet of triplets-   EDC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride-   eq. equivalent-   EtOAc ethyl acetate-   FCC flash column chromatography-   h hour-   HBTU (2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium    hexafluorophosphate-   HCl hydrochloric acid-   HOBT benztriazol-1-ol-   HPLC high pressure liquid chromatography-   HT high throughput-   H₂O water-   Hyflo Hyflo Super Cel Medium-   Isolute®SCX-2 polymer supported sulfonic acid macroporous    polystyrene-   K kelvin-   K₂CO₃ potassium carbonate-   LC liquid chromatography-   M molar-   MeCN acetonitrile-   MeOD methanol-d4-   MeOH methanol-   2-Me-THF 2-methyltetrahydrofuran-   MgS04 magnesium sulfate-   MHz mega herz-   MS mass spectroscopy-   m multiplet-   mBar millibar-   mL millilitre-   mm millimeter-   mM millimolar-   min. minute-   mw microwave-   NaOH sodium hydroxide-   Na₂SO₄ sodium sulfate-   NaHCO₃ sodium hydrogen carbonate-   NaO^(t)Bu sodium tert-butoxide-   NEt₃ triethylamine-   NH₃ ammonia-   NH₄OH concentrated solution of ammonia in water possessing a    specific gravity of 0.88-   NMP N-methylpyrrolidinone-   NMR nuclear magnetic resonance-   OBD optimum bed density-   Pd(OAc)₂ palladium acetate-   Pd(OH)₂/C palladium hydroxide on carbon-   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium-   Pd₂(dba)₃.CHCl₃ tris(dibenzylideneacetone)dipalladium chloroform    complex-   PL-HCO₃ MP polymer supported hydrogen carbonate macroporous    polystyrene-   PL-SO₃H MP polymer supported sulfonic acid macroporous polystyrene-   rt room temperature-   Rt retention time-   s singulet-   SCX-2 polymer supported sulfonic acid macroporous polystyrene-   t triplet-   TBME tert-butylmethyl ether-   tBuOK potassium tert-butoxide-   tert-BuONa sodium tert-butoxide-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   UPLC ultra performance liquid chromatography-   X-Phos dicyclohexyl(2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine

Microwave equipment used is a Biotage Initiator®

All compounds are named using AutoNom.

LCMS Methods Used:

LC Method 1 (Rt⁽¹⁾):

The retention times (Rt) were obtained on a Agilent HPLC system with anAscentis®Express column C18 30×2.1 mm, 2.7 μm (Supelco) applying agradient (H₂O+0.05% formic acid+3.75 mM Ammonium acetate)/(CH₃CN+0.04%formic acid) 90/10 to 5/95 over 3.7 min and 1.2 mL/min as solvent flowand then 5/95 over 0.7 min with 1.4 mL/min as solvent flow and 40° C.for the oven temperature. Detection method UV 220-400 nm-MS.

LC Method 2 (Rt⁽²⁾):

The retention times (Rt) were obtained on a Agilent HPLC system with anAscentis®Express column C18 30×2.1 mm, 2.7 μm (Supelco) applying agradient (H₂O+0.05% formic acid+3.75 mM Ammonium acetate)/(CH₃CN+0.04%formic acid) 95/5 to 5/95 over 3.7 min and 1.2 mL/min as solvent flowand then 5/95 over 0.7 min with 1.4 mL/min as solvent flow and 40° C.for the oven temperature. Detection method UV 220-400 nm-MS.

LC Method 3 (Rt⁽³⁾):

The retention times (Rt) were obtained on a Agilent HPLC system with anAscentis®Express column C18 30×2.1 mm, 2.7 μm (Supelco) applying agradient (H₂O+0.05% formic acid+3.75 mM Ammonium acetate)/(CH₃CN+0.04%formic acid) 99/1 over 0.5 min and 1.2 mL/min as solvent flow then 99/1to 5/95 over 1.7 min and 1.2 mL/min as solvent flow and then 5/95 over0.7 min with 1.4 mL/min as solvent flow and 40° C. for the oventemperature. Detection method UV 220-400 nm-MS.

LC Method 4 (Rt⁽⁴⁾):

The retention times (Rt) were obtained on a Agilent HPLC system with anAscentis®Express column C18 30×2.1 mm, 2.7 μm (Supelco) applying agradient (H₂O+0.05% formic acid+3.75 mM Ammonium acetate)/(CH₃CN+0.04%formic acid) 90/10 to 5/95 over 1.7 min and 1.2 mL/min as solvent flowand then 5/95 over 0.7 min with 1.4 mL/min as solvent flow and 40° C.for the oven temperature. Detection method UV 220-400 nm-MS.

LC Method 5 (Rt⁽⁵⁾):

The retention times (Rt) were obtained on a Agilent HPLC system with anAscentis®Express column C18 30×2.1 mm, 2.7 μm (Supelco) applying agradient (H₂O+0.05% TFA)/(CH₃CN+0.04% TFA) 95/5 to 5/95 over 3.7 min and1.2 mL/min as solvent flow and then 5/95 over 0.7 min with 1.4 mL/min assolvent flow and 40° C. for the oven temperature. Detection method UV220-400 nm-MS.

LC Method 6 (Rt⁽⁶⁾):

The retention times (Rt) were obtained on a Agilent HPLC system with anAscentis®Express column C18 30×2.1 mm, 2.7 μm (Supelco) applying agradient (H₂O+TFA)/(CH₃CN+0.04% TFA) 99/1 over 0.5 min and 1.2 mL/min assolvent flow then 99/1 to 5/95 over 1.7 min and 1.2 mL/min as solventflow and then 5/95 over 0.7 min with 1.4 mL/min as solvent flow and 40°C. for the oven temperature. Detection method UV 220-400 nm-MS.

LC Method 7 (Rt⁽⁷⁾):

The retention times (Rt) were obtained on a Waters Agilent HPLC systemwith an Ascentis®Express column C18 30×2.1 mm, 2.7 μm (Supelco) applyinga gradient (H₂O+0.05% TFA)/(CH₃CN+0.04% TFA) 90/10 to 5/95 over 1.7 minand 1.2 mL/min as solvent flow and then 5/95 over 0.7 min with 1.4mL/min as solvent flow and 40° C. for the oven temperature. Detectionmethod UV 220-400 nm-MS.

LC method 8 (Rt⁽⁸⁾):

The retention times (Rt) were obtained on a Waters HPLC alliance-HTsystem with an XTerra column MS C18, 50×4.6 mm, 5 μm, reverse phase,applying a gradient (H₂O+0.1% TFA)/(CH₃CN+0.1% TFA) 95/5 to 0/100 over8.0 min and 2.0 mL/min as solvent flow and 45° C. for the oventemperature. Detection method UV 220-400 nm-MS.

Purification Method:

Preparative Reverse Phase Gilson HPLC

-   -   Method A: Column SunFire prep C18 OBD 5 μm, 30×100 mm from        WATERS, with H₂O+0.1% TFA and Acetonitrile+0.1% TFA as mobile        phase. Detection method UV 220-400 nm    -   Method B: Column Atlantis prep T3 OBD 5 μm, 30×150 mm from        WATERS, with H₂O+0.1% TFA and Acetonitrile+0.1% TFA as mobile        phase. Detection method UV 220-400 nm    -   Method C: Column XTerra RP18 OBD 5 μm, 19×50 mm from WATERS,        with H₂O+0.1% TFA and Acetonitrile+0.1% TFA as mobile phase.        Detection method UV 220-400 nm

X-Ray Powder Diffraction

Instrumentation:

Method X1

Instrument Bruker AXS, D8 Advance Irradiation CuKα (30 kV, 40 mA)Detector PSD (Vantec) detector Scan range 2°-40° (2 theta value)

Method X2

Instrument Bruker D8 GADDS Discover Irradiation CuKα (40 kV, 40 mA)Detector HI-STAR Area detector Scan range 6°-40° (2 theta value)

Preparation of Intermediate Compounds

Intermediate 1: 5-Bromo-2-methoxy-3-trifluoromethyl-pyridine

To 2-methoxy-3-(trifluoromethyl)pyridine (20.0 g, 113.0 mmol) and1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (43.6 g, 152.0 mmol) wasadded TFA (80 mL) and the resulting mixture stirred at rt for 18 h underargon. The TFA was removed in vacuo (50 mbar, 45° C.) and the residuesuspended in tert-butyl methyl ether (200 mL). The resulting colourlesssolid was removed by filtration and washed with tert-butyl methyl ether(50 mL). The filtrate was concentrated in vacuo and suspended in EtOAc(50 mL) The insoluble colourless solid was removed by filtration andwashed with EtOAc (50 mL). The filtrate was concentrated in vacuo,diluted with heptane/tert-butyl methyl ether (5/1, 20 mL) and theinsoluble colourless solid was removed by filtration. The filtrate waspurified by column chromatography on silica gel with heptane/EtOAc,100/0 to 90/10. The crude product was filtered through a plug of NaHCO₃(20 g) and the filtrate evaporated in vacuo to give a golden oil (27.9g). The oil was dissolved in heptanes (20 mL) and purified by filteredthrough a plug of silica gel (80 g), eluting with heptane to give5-bromo-2-methoxy-3-(trifluoromethyl)pyridine as a colourless oil (22.5g, 74% yield). ¹H-NMR (400 MHz, DMSO-d₆, 298 K): δ ppm 4.03 (s, 3H) 7.95(d, 1H) 8.4 (d, 1H).

Intermediate 2: 1-((S)-3-Hydroxy-pyrrolidin-1-yl)-propan-1-one

(S)-Pyrrolidin-3-ol (10.0 g, 81.0 mmol), triethylamine (23.6 mL, 170.0mmol) and CH₂Cl₂ (150 mL) were combined in a pear-shaped flask to give abeige suspension. The mixture was cooled to −10° C. and propionylchloride (7.06 mL, 81.0 mmol) was added dropwise over 15 min,maintaining the temperature between −10 to 0° C. The resulting beigesuspension was stirred for 2 h at 0° C. MeOH (9.8 mL) was added and themixture allowed to warm to room temperature then stirred for 1 h to givea brown solution. The mixture was evaporated in vacuo to give a beigeresidue which was stirred in diethylether (200 mL) and filtered. Thefiltrate was evaporated in vacuo to give1-((S)-3-hydroxy-pyrrolidin-1-yl)-propan-1-one as a yellow oil (11.23 g,95% yield). ¹H-NMR (400 MHz, DMSO-d₆, 298 K): δ ppm 0.92-1.02 m, 3H)1.67-1.97 (m, 2H) 2.13-2.28 (m, 2H) 3.18-3.52 (m, 4H) 4.17-4.32 (m, 1H)4.85-4.97 (m, 1H). LCMS: [M+H]+=144.0

Intermediate 3:((S)-3-Hydroxy-pyrrolidin-1-yl)-(tetrahydro-pyran-4-yl)-methanone

The (S)-pyrrolidin-3-ol hydrochloride (3.69 g, 29.9 mmol) andtriethylamine (6.65 g, 9.16 mL, 65.7 mmol) were put in CH₂Cl₂ (15 mL).The suspension was cooled at ˜3° C. To this mixture, a solution oftetrahydro-pyran-4-carbonyl chloride (4.67 g, 29.9 mmol) in CH₂Cl₂ (15mL) was added slowly. Then the resulting reaction mixture was stirredfor 1.5 h at 3-10° C. The reaction mixture was then concentrated to givea powder. To this powder, addition of EtOAc (100 mL). The solid wasfiltered and washed with EtOAc. The recovered filtrate was thenconcentrated to give((S)-3-hydroxy-pyrrolidin-1-yl)-(tetrahydro-pyran-4-yl)-methanone asbeige powder. (6.77 g, 98% yield). ¹H-NMR (400 MHz, Methanol-d₄, 298 K):δ ppm 1.59-2.15 (m, 6H) 2.69-2.86 (m, 1H) 3.43-3.75 (m, 6H) 3.94-4.00(m, 2H) 4.37-4.48 (m, 1H). LCMS: [M+H]+=199.9, Rt⁽⁶⁾=0.86 min

Intermediate 4:[(S)-1-(Tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl]-carbamic acidtert-butyl ester

To a vigorously stirring solution of tetrahydro-2H-pyran-4-carbonylchloride (0.455 g, 3.06 mmol) in CH₂Cl₂ (10 mL) was added simultaneouslyportionwise sat. NaHCO₃(aq) (10 mL) and a solution of the(S)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (570 g, 3.06 mmol)at rt. The resulting biphasic mixture was stirred vigorously at rt for 3h. The organic layer was separated by filtration through a phaseseparation tube, concentrated in vacuo and purified by flashchromatography on silica gel with CH₂Cl₂/MeOH to give[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl]-carbamic acidtert-butyl ester as a colourless gum (0.623 g, 68% yield) LCMS:[M+H]+=299.6, Rt⁽⁷⁾=0.73 min.

Intermediate 5:(S)-3-Amino-pyrrolidin-1-yl-(tetrahydro-pyran-4-yl)-methanone

To (S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yl]-carbamic acidtert-butyl ester (intermediate 4) (0.623 g, 2.09 mmol) in CH₂Cl₂ (2.0mL) was added TFA (2.0 mL) and the resulting mixture stood at rt for 8h. Evaporated in vacuo and eluted through an Isolute® SCX-2 cartridge,eluting with methanol, then with 2M ammonia in methanol. Basic fractionswere concentrated in vacuo to give[(S)-3-amino-pyrrolidin-1-yl-(tetrahydro-pyran-4-yl)-methanone as acolourless solid (0.34 g, 82% yield) LCMS: [M+H]+=199.0, Rt⁽³⁾=0.1 min.

Intermediate 6:3-(4-Acetyl-piperazine-1-carbonyl)-1-methyl-3H-imidazol-3-ium iodide

1-(Piperazin-1-yl)ethanone (143 g, 1.12 mmol) and CDI (199 g, 1.23 mmol)were refluxed in THF (10 mL) under argon overnight. Cooled to roomtemperature, diluted with CH₂Cl₂ (20 mL) and water (5 mL) and theorganic layer filtered through a phase separation tube and concentratedin vacuo. Dissolved in acetonitrile (5 mL) in a glass vial and methyliodide (0.279 mL, 4.46 mmol) was added. The vial was capped and stood atroom temperature for 24 h. The solvent was evaporated in vacuo and theresidue triturated with heptane/EtOAc, 10/1 (10 mL) to give3-(4-acetyl-piperazine-1-carbonyl)-1-methyl-3H-imidazol-3-ium iodide asa colourless gum (400 mg) which was used without further purification.

Intermediate 7:(S)-3-(5,6,7,8-Tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

Pd(OH)₂/C (1.2 g, 1.71 mmol) was flushed with argon,(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (10.95 g, 26.7 mmol) dissolved in methanol (25 mL)was added followed by the addition of ammonium formate (1.68 g, 26.7mmol). The reaction mixture was refluxed for 1 h, cooled down to roomtemperature, filtered through a celite pad and concentrated undervacuum. Purification by flash chromatography on silica gel (CH₂Cl₂ thenTBME then TBME/MeOH 100/0 to 90/10 then TBME/MeOH/N H₄OH 85/15/5) gave(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (7.39 g, 87% yield) as a yellow sticky oil. ¹H NMR(400 MHz, methanol-d4, 298K) δ ppm 1.46-1.46 (m, 9H) 2.10-2.30 (m, 2H)2.78-2.83 (m, 2H) 3.11-3.14 (m, 2H) 3.41-3.60 (m, 3H) 3.65-3.72 (m, 1H)3.78 (s, 2H) 5.68 (m, 1H) 8.52 (s, 1H). LCMS: [M+H]⁺=321.2, Rt⁽²⁾=0.87min

Alternative Synthesis for Intermediate 7

Pd(OH)₂/C (1.54 g, 2.2 mmol) was flushed with nitrogen,(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (8.5 g, 20.67 mmol) dissolved in methanol (50 mL)was added followed by the addition of triethylammonium formate (7.9 g,53.7 mmol). The reaction mixture was refluxed for 1 h, cooled down toroom temperature, filtered through a celite pad and the filtrate waspartitioned between 2-Me-THF (50 mL) and water (20 mL). The upperorganic phase was collected and the bottom aqueous phase wasre-extracted with 2-Me-THF (10 mL). All the organic layers were combinedand concentrated under vacuum to provide(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (6.2 g, 94% yield) as a yellow gum.

(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

To a solution of (S)-3-hydroxypyrrolidine-1-carboxylic acid tert-butylester (0.94 g, 5.01 mmol) in THF (20 mL) was added under argon NaH (0.23g, 5.78 mmol). The mixture was stirred at rt for 25 min., then6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (1 g, 3.85mmol) was added and stirring continued at rt for 4 h. The mixture wasquenched with H₂O, extracted with CH₂Cl₂. The organic layer was filteredand evaporated to dryness. Purification by flash chromatography onsilica gel (heptanes/ethyl acetate, 1/1) gave the(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (1.35 g, 85% yield) as a yellow gum. ¹H NMR (400MHz, DMSO-d6, 298K) δ ppm 1.39 (s, 9H) 2.00-2.20 (m, 2H) 2.35-2.81 (m,4H) 3.36-3.63 (m, 6H) 3.70 (br. s, 2H) 5.50-5.59 (m, 1H) 7.25-7.37 (m,5H) 8.56 (s, 1H). LCMS: [M+H]+=411.6, Rt⁽⁷⁾=1.00 min

Alternative synthesis for(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester

To a solution of (S)-3-hydroxypyrrolidine-1-carboxylic acid tert-butylester (6.21 g, 33.16 mmol) and6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (9 g, 34.65mmol) in 2-Me-THF (100 mL) was added under nitrogen tBuOK (8.17 g, 72.95mmol). The mixture was stirred at rt for 25 min. The mixture wasquenched with H₂O. The organic layer was washed with brine. Theresulting organic solution was concentrated in vacuo to provide(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (12.6 g, 89% yield) as a yellow gum.

Intermediate 8:6-(2,4-Dimethoxy-pyrimidin-5-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

5-Bromo-2,4-dimethoxy-pyrimidine (89 g, 0.41 mmol), X-Phos (46 g, 0.09mmol) bis(dibenzylideneacetone)palladium(0) (29 g, 0.03 mmol), cesiumcarbonate (203 g, 0.62 mmol) were combined and flushed 10 min withArgon. To this mixture was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 7) (100 g, 0.31 mmol) in dioxane (4mL), the vial was capped and the reaction mixture was stirred at 120° C.for 4.5 h. The mixture was allowed to cool to rt and filtered through acelite pad. The filtrate was diluted with EtOAc (20 mL) and washed withsat. NaHCO₃(aq) (10 mL), brine (10 mL), dried (Na₂SO₄) and concentratedin vacuo. Dissolved in dioxane (4 mL) and added to a glass vialcontaining 5-bromo-2,4-dimethoxy-pyrimidine (89 g, 0.41 mmol), X-Phos(46 g, 0.09 mmol) tris(dibenzylideneacetone)dipalladium(0) (29 g, 0.03mmol), cesium carbonate (203 g, 0.62 mmol). The vial was capped and thereaction mixture was stirred at 120° C. for 4.5 h. The mixture wasallowed to cool to rt and filtered through a celite pad. The filtratewas diluted with EtOAc (20 mL) and washed with sat. NaHCO₃(aq) (10 mL)then brine (10 mL), dried (Na₂SO₄) and concentrated in vacuo to give(S)-3-(6-(2,4-dimethoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carboxylicacid tert-butyl ester which was used without further purification.(S)-3-(6-(2,4-dimethoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carboxylicacid tert-butyl ester was dissolved in CH₂Cl₂ (2.0 mL) and TFA added (1mL). The resulting mixture was stirred for 30 min. at room temperature.The reaction mixture was concentrated in vacuo. Purification bypreparative reverse phase Gilson HPLC and subsequent neutralization ofthe combined fractions by PL-HCO3 cartridge & lyophilisation gave6-(2,4-dimethoxy-pyrimidin-5-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineas a yellow powder (11 mg, 10% yield over 2 steps). LCMS: [M+H]+=359.1,Rt⁽²⁾=0.79 min

Intermediate 9:2-Amino-5-[4-((S)-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]nicotinonitrile

(S)-3-(5,6,7,8-Tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 7) (84 g, 0.263 mmol),imidodicarbonic acid, 2-[5-bromo-3-(cyano)-2-pyridinyl]-,1,3-bis(1,1-dimethylethyl) ester (115 g, 0.289 mmol), X-Phos (376 g,0.079 mmol), tris(dibenzylideneacetone)dipalladium(0) (24 g, 0.026mmol), cesium carbonate (171 g, 0.526 mmol) were combined in a glassvial and flushed 10 min with Argon. To this mixture was added dioxane(4.0 mL), the vial was capped and the reaction mixture stirred at 120°C. for 1.5 h. The reaction was allowed to cool to rt and filteredthrough a celite pad, The filtrate was diluted with EtOAc (20 mL) andwashed with sat. NaHCO₃(aq) (10 mL) and brine (10 mL), dried (Na₂SO₄)and concentrated in vacuo to give (S)-tert-butyl3-(6-(6-(bis(tert-butoxycarbonyl)amino)-5-(cyano)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carboxylatewhich was used without further purification. (S)-tert-butyl3-(6-(6-(bis(tert-butoxycarbonyl)amino)-5-(cyano)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carboxylatewas dissolved in CH₂Cl₂ (2.0 mL) and TFA added (1 mL). The resultingmixture was stirred for 30 min at room temperature. The reaction mixturewas concentrated in vacuo. Purification by preparative reverse phaseGilson HPLC and subsequent neutralization of the combined fractions byPL-HCO3 cartridge & lyophilisation gave2-amino-5-[4-((S)-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]nicotinonitrileas a yellow powder (17 mg, 19% yield over 2 steps). LCMS: [M+H]+=338.3,Rt⁽³⁾⁼1.16 min.

Imidodicarbonic acid, 2-[5-bromo-3-(cyano)-2-pyridinyl]-, 1,3-bis(1,1-dimethylethyl) ester

To 2-amino-5-bromonicotinonitrile (0.785 g, 3.96 mmol), triethylamine(0.553 mL, 3.96 mmol) and 4-dimethylaminoyridine (20 g, 0.164 mmol) inCH₂Cl₂ (25 mL) was added di-tert-butyl-dicarbonate (2.16 g, 9.91 mmol)and the resulting mixture stirred at room temperature for 18 h.Evaporated to dryness in vacuo and triturated in heptane (25 mL) for 72h. The resulting precipitate was filtered and washed with heptane (10mL) to give imidodicarbonic acid, 2-[5-bromo-3-(cyano)-2-pyridinyl]-,1,3-bis(1,1-dimethylethyl) ester as a beige solid (1.1 g, 70% yield). ¹HNMR (400 Mhz, CDCl₃, 298K) 1.51 (s, 18H) 8.16 (d, 1H) 8.77 (d, 1H).LCMS: [M+H]+=398/400.1, Rt⁽⁴⁾=1.43 min.

Intermediate 10:(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

To a glass vial was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 7) (1.00 g, 3.12 mmol),5-bromo-2,3-dimethoxypyridine (0.82 g, 3.75 mmol), sodium tert-butoxide(0.46 g, 4.68 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.11 g,0.13 mmol), 2-di-t-butylphosphino-2′-(N, N-dimethylamino)biphenyl (0.06g, 0.18 mmol) and anhydrous toluene (10 mL). The vial was flushed with astream of argon for 15 sec and capped. The mixture was heated withstirring for 18 h at 80° C. Allowed to cool and filtered through acelite pad. The filtrate was diluted with EtOAc (50 mL) and washed withbrine (20 mL). The organic layer was separated, dried (Na₂SO₄) andconcentrated in vacuo. Purified by flash column chromatography on silicagel with EtOAc/MeOH, 98/2 to 92/18 to give(S)-3-[6-(5,6-dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester as a pale yellow foam (1.05 g, 74% yield). LCMS:[M+H]+=458.1, Rt⁽⁴⁾=1.02 min.

Intermediate 11:(S)-3-[6-(5-Cyano-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

To a glass vial was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 7) (630 g, 1.97 mmol),5-bromo-2-methoxynicotinonitrile (419 g, 1.97 mmol), cesium carbonate(1281.0 g, 3.93 mmol), tris(dibenzylideneacetone)dipalladium(0) (180 g,0.20 mmol), X-Phos (319 g, 0.67 mmol) and anhydrous dioxane (10.0 mL).The vial was flushed with a stream of argon for 15 sec and capped. Themixture was heated with stirring for 1 h at 110° C. and then stirred atroom temperature for 18 h. Diluted with CH₂Cl₂ (100 mL) and water (30mL) and filtered through a celite pad. The organic phase was separatedby filtering through a phase separation tube and concentrated in vacuo.Purified by flash chromatography on silica gel with heptanes/EtOAc,80/20 to 0/100 to give(S)-3-[6-(5-cyano-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester as a brown gum (350 mg, 39% yield) LCMS:[M+H]+=453.6, Rt⁽⁷⁾=1.29 min.

Intermediate 12:(S)-3-[6-(5-Fluoro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

To a glass vial was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 7) (150 g, 0.47 mmol),5-bromo-3-fluoro-2-methoxypyridine (96 g, 0.47 mmol), cesium carbonate(305 g, 0.94 mmol), tris(dibenzylideneacetone)dipalladium(0) (43 g, 0.05mmol), X-Phos (76 g, 0.16 mmol) and anhydrous dioxane (2.0 mL). The vialwas flushed with a stream of argon for 15 sec and capped. The mixturewas heated with stirring for 1.5 h at 110° C. and then stirred at roomtemperature for 18 h. Diluted with CH₂Cl₂ (25 mL), filtered through acelite pad and concentrated in vacuo. Purified by reverse phase GilsonHPLC (Method A) to give(S)-3-[6-(5-fluoro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester trifluoroacetate as a brown gum (45 mg, 17% yield)LCMS: [M+H]+=446.4, Rt⁽⁴⁾=1.41 min.

Intermediate 13:(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

To a glass vial was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 7) (150 g, 0.47 mmol)5-bromo-2-methoxy-3-(trifluoromethyl)pyridine (intermediate 1)(120 g,0.47 mmol), cesium carbonate (305 g, 0.94 mmol),tris(dibenzylideneacetone)dipalladium(0) (43 mg, 0.05 mmol), X-Phos (76g, 0.16 mmol) and anhydrous dioxane (2.0 mL). The vial was flushed witha stream of argon for 15 sec and capped. The mixture was heated withstirring for 1 h at 110° C. and then stirred at room temperature for 18h. Diluted with CH₂Cl₂ (10 mL) and water (2 mL), filtered through acelite pad. The organic phase was separated by filtering through a phaseseparation tube and concentrated in vacuo. Purified by reverse phaseGilson HPLC (Method A) to give (S)-tert-butyl3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carboxylatetrifluoroacetate(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester trifluoroacetate as a brown gum (90 mg, 32% yield)LCMS: [M+H]+=496.5, Rt⁽⁷⁾=1.43 min.

Intermediate 14:4-Methoxy-6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

To a glass vial was added4-methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (WO 2008/130481, p47) (0.570 g, 3.45 mmol), 5-bromo-2-methoxy-3-methylpyridine (0.697 g,3.45 mmol), cesium carbonate (2.25 g, 6.90 mmol),tris(dibenzylideneacetone)dipalladium(0) (0.316 g, 0.345 mmol), X-Phos(0.493 g, 1.04 mmol) and anhydrous dioxane (5 mL). The vial was flushedwith a stream of argon for 15 sec and capped. The mixture was heatedwith stirring for 1 h 45 min at 110° C. then allowed to cool to roomtemperature and stirred at RT for 3 days. The reaction mixture wasfiltered through a celite pad and concentrated in vacuo.

Purified by flash chromatography on silica gel with heptane/EtOAc, 100/0to 0/100 then EtOAc/MeOH, 90/10 to give4-methoxy-6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineas a brown gum (0.36 g, 36% yield) LCMS: [M+H]+=287.0, Rt⁽⁷⁾=0.80 min.

Intermediate 15:6-(5-Chloro-6-methoxy-pyridin-3-yl)-4-methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

To a glass vial was added4-methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (WO 2008/130481, p47) (0.273 g, 1.65 mmol), 5-bromo-3-chloro-2-methoxypyridine (0.368 g,1.65 mmol), sodium tert-butoxide (318 g, 3.31 mmol), diacetoxypalladium(0.037 g, 0.17 mmol), X-Phos (0.079 g, 0.17 mmol) and anhydroustoluene/tert-butanol, 5/1 (6 mL). The vial was flushed with a stream ofargon for 15 sec and capped. The mixture was heated with stirring for 2h at 110° C. then allowed to cool to room temperature and stirred at rtfor 5 days. Diluted with CH₂Cl₂ (10 mL) and water (2 mL), filteredthrough a celite pad. The organic phase was separated by filteringthrough a phase separation tube and concentrated in vacuo. Purified byflash chromatography on silica gel with heptane/EtOAc 100/0 to 0/100 togive6-(5-chloro-6-methoxy-pyridin-3-yl)-4-methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineas a yellow solid (95 mg, 19% yield) LCMS: [M+H]+=307.0/308.9,Rt⁽³⁾=1.62 min.

Intermediate 16:4-Methoxy-6-(5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

To a glass vial was added4-methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (0.273 g, 1.65mmol), 3-bromo-5-(trifluoromethyl)pyridine (0.373 g, 1.65 mmol), cesiumcarbonate (1.08 g, 3.31 mmol), tris(dibenzylideneacetone)dipalladium(0)(0.076 g, 0.083 mmol), X-Phos (0.079 g, 0.165 mmol) and anhydrousdioxane (5 mL). The vial was flushed with a stream of argon for 15 secand capped. The mixture was heated with stirring for 1.5 h at 110° C.Filtered through a celite pad, concentrated in vacuo and purified byflash chromatography on silica gel with heptanes/EtOAc, 100/0 to 0/100to give4-methoxy-6-(5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineas an orange gum (195 mg, 34% yield)¹H NMR (DMSO-d6, 298K) 2.95 (t, 2H)3.77 (t, 2H) 4.02 (s, 3H) 4.37 (s, 2H) 7.67-7.71 (m, 1H) 8.30-8.34 (m,1H) 8.63 (s, 1H) 8.67-8.71 (1H, m) LCMS: [M+H]+=311.2, Rt⁽⁴⁾=0.94 min.

Intermediate 17:6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ol

To4-methoxy-6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(intermediate 14)(360 g, 1.26 mmol) in MeOH (2.0 mL) in a glass vial wasadded 2M NaOH(aq) (2.0 mL). The vial was capped and heated at 90° C. for24 h. Acidified with glacial AcOH to pH 6, evaporated in vacuo and theresidue extracted with CH₂Cl₂ (2×30 mL). With each extraction, theCH₂Cl₂ layer was decanted from the solid residue. The CH₂Cl₂ layers werecombined and eluted through an Isolute®SCX-2 cartridge, eluting withmethanol, then with 2M ammonia in methanol. Basic fractions wereconcentrated in vacuo to give6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-olas a brown gum (260 mg, 76% yield) LCMS: [M+H]+=273.1, Rt⁽³⁾=1.33 min.

Intermediate 18:6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ol

To6-(5-chloro-6-methoxy-pyridin-3-yl)-4-methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(intermediate 15)(95 g, 0.31 mmol) in MeOH (5.0 mL) in a glass vial wasadded 2M NaOH(aq) (3.0 mL). The vial was capped and heated at 90° C. for24 h. Acidified with glacial AcOH to pH 6, evaporated in vacuo and theresidue extracted with CH₂Cl₂ (1×50 mL with stirring). With eachextraction, the CH₂Cl₂ layer was decanted from the solid residue. TheCH₂Cl₂ layers were combined. The solid residue was then washed withwater (10 mL) and filtered. This filtered solid was combined with theCH₂Cl₂ layers and evaporated in vacuo to give6-(5-chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-olas a yellow solid (90 mg, 107% yield) LCMS: [M+H]+=293.0/294.8,Rt⁽³⁾=1.38 min.

Intermediate 19:6-(5-Trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-01

To4-methoxy-6-(5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(intermediate 16) (190 g, 0.612 mmol) in MeOH (2.0 mL) in a glass vialwas added 2M NaOH(aq) (2.0 mL). The vial was capped and heated at 90° C.for 24 h. Acidified with glacial AcOH to pH 6, evaporated in vacuo andthe residue extracted with CH₂Cl₂ (2×30 mL with sonication). With eachextraction, the CH₂Cl₂ layer was decanted from the solid residue. TheCH₂Cl₂ layers were combined and eluted through an Isolute® SCX-2cartridge, eluting with methanol, then with 2M ammonia in methanol.Basic fractions were concentrated in vacuo to give6-(5-(trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-olas a yellow solid (167 mg) LCMS: [M+H]+=297.2, Rt⁽⁴⁾=0.69 min.

Intermediate 20:(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester

To6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ol(intermediate 17) (178 g, 0.654 mmol) in acetonitrile (2.0 mL) was addedBOP (376 g, 0.854 mmol) and DBU (0.197 mL, 1.31 mmol). The resultingsolution was stood at rt for 2 min then added (S)-tert-butyl3-aminopyrrolidine-1-carboxylate (365 g, 1.96 mmol) in acetonitrile (2.0mL) and heated the mixture at 75° C. for 72 h. The reaction mixture wasevaporated in vacuo and purified by reverse phase Gilson HPLC (Method A)to give(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester trifluoroacetate (60 mg, 17% yield) as a browngum. LCMS: [M+H]+=441.2, Rt⁽³⁾=1.50 min

Intermediate 21:(S)-3-[6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester

To6-(5-chloro-6-methoxypyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ol(intermediate 18) (90 g, 0.31 mmol) in acetonitrile (3.0 mL) was addedBOP (177 mg, 0.40 mmol) and DBU (0.15 mL, 0.99 mmol). The resultingsolution was stood at rt for 2 min then added (S)-tert-butyl3-aminopyrrolidine-1-carboxylate (0.17, 0.93 mmol) and heated themixture at 70° C. for 96 h. The reaction mixture was evaporated in vacuoand purified by reverse phase Gilson HPLC (Method A) to give(S)-3-[6-(5-chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester trifluoroacetate (50 mg, 35% yield) as a browngum. LCMS: [M+H]+=461.1/463.0, Rt (4)=0.93 min.

Intermediate 22:(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester

6-Benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (5.0 g,19.06 mmol), (S)-tert-butyl 3-aminopyrrolidine-1-carboxylate (4.11 g,20.96 g) and triethylamine (3.98 mL, 28.6 mmol) were heated in a sealedvial at 120° C. for 42 h. The mixture was allowed to cool, diluted withtert-butyl methyl ether (100 mL) and the resulting suspension stirredfor 10 min. The mixture was diluted with water (50 mL) and the organiclayer separated. The organic layer was washed with brine (20 mL), dried(Na₂SO₄) and evaporated in vacuo to give a brown gum. The residue waspurified by column chromatography on silica gel with EtOAc/MeOH, 98/2 to82/18 to give(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester as a pale yellow foam (7.36 g, 93% yield). ¹H-NMR(400 MHz, CDCl₃, 298 K): δ ppm 1.48 (s, 9H) 2.10-2.31 (m, 2H) 2.80-2.96(m, 4H) 3.15-3.87 (m, 8H) 4.44-4.77 (m, 1H) 5.62-5.73 (m, 1H) 7.29-7.45(m, 5H) 8.50 (s, 1H). LCMS: [M+H]+=410.0, Rt⁽⁶⁾=1.39 min.

Alternative Synthesis for Intermediate 22

(S)-tert-Butyl-3-aminopyrrolidine-1-carboxylate (50 g, 192.5 mmol) wasadded to 6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(39.440 g, 211.8 mmol) in NMP (200 mL) solution followed by the additionof K₂CO₃ (39.9 g, 288.8 mmol). The mixture was heated to 120° C. for 20h. The mixture was allowed to cool, partitioned between water (300 mL)and ethylacetate (500 mL). the bottom aqueous phase was discarded andthe upper organic phase was washed with brine (150 mL) and concentratedin vacuo to provide crude(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester as a pale yellow foam (76.44 g, 97% yield).

Intermediate 23:(S)-3-(5,6,7,8-Tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester

To a solution of(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 22) (30.1 g, 73.5 mmol) in MeOH (100mL) was added 20% palladium hydroxide on carbon (3.3 g) then ammoniumformate (4.63 g, 73.5 mmol) and the mixture heated at reflux for 1 h.Added ammonium formate (0.38 g, 6.02 mmol) and continued heating atreflux for 30 min. The reaction mixture was allowed to cool and filteredthrough a celite pad, washing with MeOH (50 mL) then CH₂Cl₂ (50 mL). Thefiltrate was evaporated in vacuo to give a brown oil. Dissolved inCH₂Cl₂ (100 mL), added solid NaHCO₃ (10 g) and filtered through a celitepad. The filtrate was evaporated in vacuo to give a brown oil. Dissolvedin EtOAc (50 mL) and a solid precipitated which was filtered to give(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester as a beige solid (15.55 g, 66% yield). ¹H-NMR (400MHz, DMSO-d₆, 298 K): δ ppm 1.40 (s, 9H) 1.81-1.98 (m, 1H) 2.05-2.17 (m,1H) 2.92 (t, 2H) 3.10-3.46 (m, 5H) 3.49-3.63 (m, 3H) 4.47-4.63 (m, 1H)6.46 (d, 1H, N—H) 8.25 (s, 1H). LCMS: [M+H]+=320.0, Rt⁽⁶⁾=1.29 min.

Alternative Synthesis for Intermediate 23

Pd(OH)₂/C (6.60 g, 5.3 mmol) was flushed with nitrogen,(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 22) dissolved in methanol (164 mL)was added followed by the addition of triethylammonium formate (28.4 g,188.0 mmol). The reaction mixture was refluxed for 1 h, cooled down toroom temperature, filtered through a celite pad and the filtrate wasconcentrated under vacuum. the residue was recrystallized with methyltert-butyl ether (200 mL) and heptanes (50 mL) to provide(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester as a beige solid (25.7 g, 85% yield).

Intermediate 24:(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester

To a glass vial was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 23) (3.5 g, 10.96 mmol),5-bromo-2-methoxy-3-(trifluoromethyl)pyridine (intermediate 1) (3.09 g,12.05 mmol), sodium tert-butoxide (1.58 g, 16.44 mmol),tris(dibenzylideneacetone)dipalladium(0) (0.502 g, 0.548 mmol),2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl (0.225 g, 0.657mmol) and anhydrous tert-butanol (6 mL). The vial was flushed with astream of argon for 15 sec and capped. The mixture was heated withstirring for 5 h at 100° C. Allowed to cool and partitioned betweenEtOAc (100 mL) and water (20 mL) and filtered the biphasic mixturethrough a celite pad. The organic layer was separated, dried (MgSO₄) andconcentrated in vacuo. Purified by flash column chromatography throughBiotage® amino silica gel eluting with heptane/EtOAc, 100/0 to 0/100then EtOAc/MeOH (90/10) to give(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester as a yellow foam (4.00 g, 74% yield). LCMS:[M+H]+=495.2, Rt⁽³⁾=1.59 min.

Alternative Synthesis for Intermediate 24

To a glass flask was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 23) (6.331 g, 15.86 mmol),5-bromo-2-methoxy-3-(trifluoromethyl)pyridine (intermediate 1) (4.465 g,17.442 mmol), sodium tert-butoxide (2.29 g, 23.78 mmol),tris(dibenzylideneacetone)dipalladium(0) (0.726 g, 0.793 mmol),di-tert-butyl(2′-methylbiphenyl-2-yl)phosphine (0.297 g, 0.951 mmol) andanhydrous tert-butanol (30 mL). The flask was flushed with a stream ofnitrogen for 15 sec and capped. The mixture was heated with stirring for4 h under reflux. The mixture was allowed to cool to rt and partitionedbetween EtOAc (100 mL) and water (20 mL). The biphasic mixture wasfiltered the through a celite pad. The organic layer was separated andconcentrated in vacuo to give crude(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester as a yellow foam (7.46 g, 95% yield).

Intermediate 25:(S)-3-[6-(5-Cyano-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester

To a glass vial was added(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 23) (566 g, 1.77 mmol),5-bromo-2-methoxynicotinonitrile (453 g, 2.13 mmol), cesium carbonate(1155 mg, 3.54 mmol), tris(dibenzylideneacetone)dipalladium(0) (162 g,0.18 mmol), X-Phos (287 mg, 0.60 mmol) and anhydrous tert-butanol (5mL). The vial was flushed with a stream of argon for 15 sec and capped.The mixture was heated with stirring for 18 h at 110° C. Allowed to cooland partitioned between CH₂Cl₂ (20 mL) and water (10 mL) and filteredthe biphasic mixture through a celite pad. The organic layer wasseparated by filtering through a phase separation tube and concentratedin vacuo. Purified by flash column chromatography on silica gel withheptane/EtOAc, 100/0 to 0/100 then EtOAc/MeOH (90/10) to give(S)-3-[6-(5-cyano-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester as a brown gum (234 mg, 29% yield). LCMS:[M+H]+=452.1, Rt⁽⁴⁾=0.90 min.

Preparation of Examples

a)(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester III is firstly prepared by reacting6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine with(S)-3-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester in thepresence of a suitable base such as sodium hydride (NaH) and polarorganic solvent such as THF or dioxane under inert gas conditions atroom temperature. b) N-debenzylation is performed under customarytransfer hydrogenation conditions, using among the possible palladiumcatalysts, preferably palladium hydroxide on carbon Pd(OH)₂/C and amongthe possible formate salt preferably ammonium formate and organicsolvent such as preferably methanol. The reaction is preferably carriedout under refluxing conditions. c) Buchwald-Hartwig cross couplingbetween(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester IV and aryl bromide of the general formula R²—Xwhere X=Bromo or Iodo is performed under customary Buchwald-Hartwigconditions using such a ligand such as X-Phos or2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl with a palladiumcatalyst such as Pd₂(dba)₃ or Pd₂(dba)₃.CHCl₃ or Pd(OAc)₂, preferablyPd₂(dba)₃ with X-Phos, base such as preferably Cs₂CO₃ or preferablytert-BuONa, and organic solvent such as preferably dioxane or preferablyTHF. The reaction is preferably stirred at a temperature ofapproximately 80-120° C., preferably 120° C. The reaction may preferablycarry out under an inert gas such as nitrogen or argon. d) N-BOCdeprotection is performed under customary BOC deprotection conditionsusing among the possible acid preferably trifluoro-acetic acid or HCland suitable organic solvent such as CH₂Cl₂ or diethyl ether. Thereaction is preferably performed at room temperature. e)

Reaction of compounds of general formula VI with an acid chloride offormula R⁴C(O)Cl or carboxylic acid of formula R⁴C(O)OH. Those skilledin the art will appreciate that there are many known ways of preparingamides. For example, see Mantalbetti, C. A. G. N and Falque, V., Amidebond formation and peptide coupling, Tetrahedron, 2005, 61(46), pp10827-10852 and references cited therein. The examples provided hereinare thus not intended to be exhaustive, but merely illustrative.

The following general methods i-v have been used.

i. To a vigorously stirring solution of the acid chloride (1.3 eq.) inCH₂Cl₂ was added simultaneously portionwise excess sat. NaHCO₃(aq) and asolution of the amine of general formula VI (1.0 eq.) in CH₂Cl₂ at rt.The resulting biphasic mixture was stirred vigorously at rt for 2 h. Theorganic layer was separated, dried (MgSO₄), concentrated in vacuo andpurified by either reverse phase chromatography, normal phasechromatography or crystallisation.

ii. To the amine of general formula VI (1.0 eq.) in CH₂Cl₂ was added theacid chloride (1.1 eq.) and triethylamine (3.0 eq.) at rt. The reactionmixture was stirred at room temperature for 1 h. The reaction mixturewas concentrated under vacuum and subsequently partitioned between waterand a suitable organic solvent and purified either reverse phasechromatography, normal phase chromatography or crystallisation.

iii. To the carboxylic acid (1.0 eq.) and HBTU (1.2 eq.) in DMF wasadded triethylamine (4.0 eq.). The mixture was stirred for 20 min andthen the amine of general formula VI (1.0 eq.) in DMF was added. Themixture was allowed to stir overnight at room temperature andsubsequently partitioned between water and a suitable organic solvent.The organic phase was separated, dried (MgSO₄), concentrated in vacuoand purified by either reverse phase chromatography, normal phasechromatography or crystallisation.

iv. To the carboxylic acid (1.0 eq.) and the amine general formula VI(1.0 eq.) in DMF was added DCC (1.2 eq.) in DMF. The reaction mixturewas stirred at rt for 18 h and concentrated in vacuo and purified byeither reverse phase chromatography, normal phase chromatography orcrystallisation.

v. To the carboxylic acid (1.1 eq.) and the amine general formula VI(1.0 eq.) in CH₂Cl₂ was added benztriazol-1-ol (1.1 eq.) and EDC (1.6eq.). The reaction mixture was stirred at rt for 18 h and subsequentlypartitioned between water and a suitable organic solvent. The organicphase was separated, dried (MgSO₄), concentrated in vacuo and purifiedby either reverse phase chromatography, normal phase chromatography orcrystallization.

a) N-BOC deprotection is performed under customary BOC deprotectionconditions using among the possible acid preferably trifluoro-aceticacid and organic solvent preferably CH₂Cl₂. The reaction is preferablyperformed at room temperature. b) Reaction of compound of generalformula IX with an acid chloride of formula R⁴C(O)Cl or carboxylic acidof formula R⁴C(O)OH using general methods i-v as described in Scheme 1,step e. Those skilled in the art will appreciate that there are manyknown ways of preparing amides. For example, see Mantalbetti, C. A. G. Nand Falque, V., Amide bond formation and peptide coupling, Tetrahedron,2005, 61(46), pp 10827-10852 and references cited therein. The examplesprovided herein are thus not intended to be exhaustive, but merelyillustrative. c) Removal of the benzyl protecting group is performedusing standard methodology as described in “Protecting groups in OrganicSynthesis” by T. W. Greene and P. Wutz, 3^(rd) edition, 1999, John Wileyand Sons. Typical conditions comprise of 1.0 eq. of compound of generalformula X (8.0 eq. of ammonium formate and 20% (w/w) palladium hydroxidePd(OH)₂/C (catalyst) heated under reflux in methanol. d)Buchwald-Hartwig cross coupling between compound of general formula XIand compounds of general formula R²—X where X=Bromo or Iodo is performedunder customary Buchwald-Hartwig conditions using such a ligand such asX-Phos or 2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl with apalladium catalyst such as Pd₂(dba)₃ or Pd₂(dba)₃.CHCl₃ or Pd(OAc)₂,preferably Pd₂(dba)₃ with X-Phos, base such as preferably Cs₂CO₃ orpreferably tert-BuONa, and organic solvent such as preferably dioxane orpreferably THF. The reaction is preferably stirred at a temperature ofapproximately 80-150° C., preferably 120° C. The reaction may preferablybe carried out under an inert gas such as nitrogen or argon.

Compounds of general formula XVII can be prepared in a similar manner asdescribed for steps a-e in Scheme 1, starting from6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (I) andtert-butyl 3-hydroxyazetidine-1-carboxylate (XII).

a)(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester XIX is firstly prepared by reacting6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine with(S)-3-Amino-pyrrolidine-1-carboxylic acid tert-butyl ester in thepresence of a suitable base such as triethylamine orN,N-diisopropylethylamine at elevated temperature (e.g. 120° C.) for24-48 h. Typical conditions comprise of 1.0 eq. of6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine, 1.0 eq. of(S)-3-amino-pyrrolidine-1-carboxylic acid tert-butyl ester and 1.5 eq.of triethylamine at 120° C. for 48 h. b) Removal of the benzylprotecting group is performed using standard methodology as described in“Protecting groups in Organic Synthesis” by T. W. Greene and P. Wutz,3^(rd) edition, 1999, John Wiley and Sons. Typical conditions compriseof 1.0 eq. of(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester XIX, 1.1-8.0 eq. of ammonium formate and 20% (w/w)palladium hydroxide Pd(OH)₂/C (catalyst) heated under reflux inmethanol. c)(S)-3-(5,6,7,8-Tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester XX is reacted with halide R²—X (where R2 isdefined above and X is halo and preferably bromo or iodo), in thepresence of a suitable base such as sodium tert-butoxide or cesiumcarbonate and a suitable catalyst system such as Pd₂(dba)₃ with2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl or Pd₂(dba)₃ withX-Phos in a suitable solvent such as anhydrous tert-butanol or anhydrousdioxane, heated at elevated temperature (e.g. 100° C.). The reaction maypreferably be carried out under an inert gas such as nitrogen or argon.Typical conditions comprise of 1 eq. of(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester XX, 1-1.5 eq. of R2-X, 1.5-2.0 eq. of sodiumtert-butoxide, 5-10 mol % Pd₂(dba)₃ and 5-10 mol %2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl in anhydroustert-butanol at 100° C. for 5-24 hours under an atmosphere of argon. d)N-Boc deprotection is performed under customary Boc deprotectionconditions with a suitable acid such as trifluoroacetic acid in asuitable solvent such as CH₂Cl₂ at room temperature. Typical conditionscomprise of 1 eq. of compound of general formula XII in excesstrifluoroacetic acid in CH₂Cl₂ at room temperature for 1-3 h. e)Reaction of compounds of general formula XXII with an acid chloride offormula R⁴C(O)Cl or carboxylic acid of formula R⁴C(O)OH using generalmethods i-v as described in Scheme 1, step e. Those skilled in the artwill appreciate that there are many known ways of preparing amides. Forexample, see Mantalbetti, C. A. G. N and Falque, V., Amide bondformation and peptide coupling, Tetrahedron, 2005, 61(46), pp10827-10852 and references cited therein. The examples provided hereinare thus not intended to be exhaustive, but merely illustrative.

a) 4-Methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (WO 2008/130481,p 47) is reacted with halide R²—X (where R2 is defined above and X ishalo and preferably bromo or iodo), in the presence of a suitable basesuch as cesium carbonate or sodium tert-butoxide and a suitable catalystsystem such as Pd₂(dba)₃ with X-Phos or Pd(OAc)₂ with X-Phos in asuitable solvent such as dioxane or THF, heated at elevated temperature(e.g. 110° C.). The reaction may preferably be carried out under aninert gas such as nitrogen or argon. Typical conditions comprise of 1eq. of 4-methoxy-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine, 1-1.5 eq.of R2-X, 1.5-2.0 eq. of cesium carbonate, 5-10 mol % Pd₂(dba)₃ and 5-10mol % X-Phos in dioxane at 110° C. for 5-24 hours under an atmosphere ofargon. b) Compounds of general formula XXV is reacted with aqueoussodium hydroxide in a suitable solvent such as methanol or dioxane atelevated temperature (e.g. 100° C.) for 18-24 h. Typical conditionscomprise of 1 eq. of compounds of general formula XXV in excess 2Nsodium hydroxide(aq) in methanol at 100° C. for 18 h. c) Compounds ofgeneral formula XXI can be prepared using a base promoted phosphoniumcoupling reaction whereby compounds of general formula XXVI in asuitable solvent such as acetonitrile is reacted with a phosphonium saltsuch as benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (BOP) in the presence of a base such as1,8-diaza-7-bicyclo[5.4.0]undecene (DBU) followed by addition of(S)-tert-butyl 3-aminopyrrolidine-1-carboxylate. The reaction mixture ispreferably stirred at a temperature of 20° C. to 90° C. for 18-72 h. Thereaction may preferably be carried out under an inert gas, e.g. nitrogenor argon. Typical conditions comprise of 1 equivalent of compounds ofgeneral formula XXVI, 1.0-1.5 eq. of BOP, 2.0-4.0 eq. of DBU and 2.0-3.0eq. of (S)-tert-butyl 3-aminopyrrolidine-1-carboxylate in acetonitrileat 65° C. for 72 hours under argon. Steps d) and e) can be carried outin a similar manner described for steps d) and e) in Scheme 1. Step f)can be carried out using a base promoted phosphonium coupling reactionin a similar manner as step c) in Scheme 5. Typical conditions compriseof 1 eq. of compounds of general formula XXVI, 1.0-1.5 eq. of BOP,2.0-4.0 eq. of DBU and 2.0-3.0 eq. of amine of general formula XVII inacetonitrile at 90° C. for 24 hours under argon.

a) Alcohol of general formula XXVIII is reacted with the6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine undercustomary conditions by deprotonation of the secondary alcohol usingsodium hydride (NaH) and organic solvent THF under inert gas conditionsat room temperature. b) N-debenzylation is performed under customarytransfer hydrogenation conditions, using among the possible palladiumcatalysts, preferably palladium hydroxide Pd(OH)₂ and among the possibleformate salt preferably ammonium formate and organic solvent such aspreferably methanol. The reaction is preferably carried out underrefluxing conditions. c) Buchwald-Hartwig cross coupling betweencompound of general formula XI and compounds of general formula R²—X isperformed under customary Buchwald-Hartwig conditions using such aligand such as X-Phos or2-di-t-butylphosphino-2′-(N,N-dimethylamino)biphenyl with a palladiumcatalyst such as Pd₂(dba)₃ or Pd₂(dba)₃.CHCl₃ or Pd(OAc)₂, preferablyPd₂(dba)₃ with X-Phos, base such as preferably Cs₂CO₃ or preferablytert-BuONa, and organic solvent such as preferably dioxane or preferablyTHF. The reaction is preferably stirred at a temperature ofapproximately 80-150° C., preferably 120° C. The reaction may preferablycarried out under an inert gas such as nitrogen or argon.

a) Compounds of general formula VI is reacted with phosgene in asuitable solvent such as CH₂Cl₂ in the presence of a suitable base suchas triethylamine or N,N-diisopropylethylamine at a temperature of 00° C.to 25° C. for 1-2 hours. The reaction may preferably be carried outunder an inert gas such as nitrogen or argon. Typical conditionscomprise 1.0 eq. of compound of general formula VI, 1.0-5.0 eq. ofphosgene, 3.0-4.0 eq. of triethylamine in CH₂Cl₂ under argon for 1 hour.b) Compound of general formula XXIX is reacted with amine R5R6NH in thepresence of a suitable base such as triethylamine orN,N-diisopropylethylamine in a suitable solvent such as CH₂Cl₂ orN,N-dimethylformamide at a temperature of 10° C. to 30° C. for 1-18 h.The reaction may preferably be carried out under an inert gas such asnitrogen or argon. Typical conditions comprise 1.0 eq. of compound ofgeneral formula XXIX, 1.0-1.2 eq. of R⁵R6NH, 3.0-4.0 eq. oftriethylamine in CH₂Cl₂ under argon for 2 hours. c) Compounds of generalformula VI is reacted with carbamoyl chloride R⁵R⁶NCOCl in the presenceof a suitable base such as triethylamine or N,N-diisopropylethylamine ina suitable solvent such as CH₂Cl₂ or N,N-dimethylformamide at atemperature of 0° C. to 25° C. for 1-18 hours. The reaction maypreferably be carried out under an inert gas such as nitrogen or argon.Typical conditions comprise 1.0 eq. of compound of general formula VI,1.0-1.2 eq. of R⁵R⁶NCOCl, 3.0-4.0 eq. of triethylamine in CH₂Cl₂ underargon for 18 hours. d) Compounds of general formula VI is reacted withcompounds of general formula XXXI in the presence of a suitable basesuch as triethylamine or N,N-diisopropylethylamine in a suitable solventsuch as CH₂Cl₂ or N,N-dimethylformamide at a temperature of 0° C. to 25°C. for 1-18 hours. The reaction may preferably be carried out under aninert gas such as nitrogen or argon. Typical conditions comprise 1.0 eq.of compound of general formula VI, 1.0-1.2 eq. of compound of generalformula XXXI, 1.0-2.0 eq. of triethylamine in CH₂Cl₂ under argon for 18hours. e) Compounds of general formula VI is reacted with compounds offormula R⁷OCOCl in the presence of a suitable base such as triethylamineor N,N-diisopropylethylamine in a suitable solvent such as CH₂Cl₂ orN,N-dimethylformamide at a temperature of 0° C. to 25° C. for 1-18hours. The reaction may preferably be carried out under an inert gassuch as nitrogen or argon. Typical conditions comprise 1.0 eq. ofcompound of general formula VI, 1.0-1.2 eq. of compounds of generalformula R⁷OCOCl, 3.0-4.0 eq. of triethylamine in CH₂Cl₂ under argon for18 hours.

a) Quaternarization of the tertiary amine of general formula XXXIII(where R⁸=alkyl e.g. benzyl) with compound of general formula R⁹—X(where R⁹=alkyl e.g. methyl and X=Bromo or Iodo) under customaryconditions using in particular acetone as organic solvent. b) Alkylationof amine of general formula R²—NH2 with quaternary amine XXXIV wasperformed by using base such a in particular K₂CO₃ and organic solventsuch as in particular a 2/1 mixture of ethanol and water and heating thereaction mixture at 80-100° C., in particular 80° C. c) Compound ofgeneral formula XXXV was reacted with base such as in particular NaH andcompound of general formula (R¹⁰O)₂CO (where R¹⁰=alkyl e.g. carbonicacid dimethyl ester). The reaction mixture is stirred under hightemperature (90° C.). d) Pyrimidine ring formation was obtained byreacting the compound of general formula XXXVI with formamidine acetatewith a base such as sodium methoxide and organic solvent such asmethanol at elevated temperature such as 90° C. for 2-18 h. e) Compoundof general formula XXVI was reacted with phosphoryl chloride in presenceof base such as triethylamine in organic solvent such as toluene atelevated temperature such as 100° C. for 12-18 h. f) Alcohol of generalformula XXVIII is reacted with Compound of general formula XXXVII undercustomary conditions by deprotonation of the secondary alcohol usingsodium hydride (NaH) and organic solvent THF under inert gas conditionsat room temperature.

Where it is stated that compounds were prepared in the manner describedfor an earlier example, the skilled person will appreciate that reactiontimes, number of equivalents of reagents and reaction temperatures maybe modified for each specific reaction, and that it may nevertheless benecessary or desirable to employ different work-up or purificationconditions.

Example 1:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanoneSynthesis of Example 1—Method 1a (According to Scheme 8)

Sodium hydride (60% in dispersion oil, 17.88 g, 0.447 mmol) was addedunder argon to a solution of intermediate 3 (75 g, 0.378 mmol) in 2 mLof dry THF. The suspension was stirred under an atmosphere of argon atambient temperature for 15 min.4-Chloro-6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(100 mg, 0.344 mmol) was added and stirred at rt for an additional 3hours. The reaction mixture was quenched with H₂O, extracted withCH₂Cl₂. The organic layer was washed with brine, dried over Na₂SO₄,filtered and evaporated to dryness. Purification by flash-chromatographyon silica gel (CH₂Cl₂/MeOH 95/5) gave{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanoneas a light yellow gum (115 mg, 74% yield). ¹H-NMR (400 MHz, methanol-d4,298K) δ ppm 1.59-1.87 (m, 4H) 2.20 (s, 3H) 2.27-2.43 (m, 2H) 2.74-2.91(m, 1H) 2.97-3.03 (m, 2H) 3.42-4.14 (m, 15H) 5.75-5.86 (m, 1H) 7.39-7.43(m, 1H) 7.63-7.68 (m, 1H) 8.57-8.61 (m, 1H). LCMS: [M+H]⁺=454.2,Rt⁽³⁾=1.46 min.

4-Chloro-6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

A mixture of6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ol(650 g, 2.387 mmol), phosphoroxy chloride (0.334 mL, 3.58 mmol),triethylamine (0.665 mL, 4.77 mmol) and toluene (12 mL) was heated at100° C. for 16 h. The mixture was neutralized with the addition of solidsodium bicarbonate, filtered and the solution was concentrated invacuum. The remaining black residue was taken up in CH₂Cl₂ and water,the layers were separated and the organic phase washed with brine, driedover sodium sulfate, filtered and concentrated to give a dark brownsolid. The solid was triturated in ethylacetate, filtered and driedunder high vacuum to yield4-chloro-6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(630 mg, 91% yield) as a tan solid. ¹H-NMR (400 MHz, DMSO-d6, 298K) δppm 2.15 (s, 3H) 3.03 (t, 2H) 3.53 (t, 2H) 3.82 (s, 3H) 4.26 (s, 2H)7.49 (dd, 1H) 7.74 (d, 1H) 8.85 (s, 1H). LCMS: [M+H]⁺=291.1, Rt⁽⁴⁾=0.97min.

6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ol

A mixture of6′-methoxy-5′-methyl-4-oxo-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-3-carboxylicacid methyl ester (900 g, 3.23 mmol), formamidine acetate (521 g, 4.85mmol), sodium methoxide (5.4 Molar) in methanol (2.395 mL, 12.94 mmol)and methanol (4 mL) was heated to 90° C. for 3 h. The mixture wasallowed to cool down to rt, diluted in CH₂Cl₂, neutralized with aceticacid (0.741 mL, 12.94 mmol) and quenched with H₂O. The layers wereseparated and aqueous was washed twice with CH₂Cl₂, organics werecombined, washed with brine, dried over sodium sulfate, filtered andevaporated to give a yellow solid. The solid was triturated inethylacetate to yield6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ol(669 mg, yield 76%) as a white powder. ¹H-NMR (400 MHz, DMSO-d6, 298K) δppm 2.14 (s, 3H) 2.72 (t, 2H) 3.39 (t, 2H) 3.81 (s, 3H) 3.90 (s, 2H)7.42 (d, 1H) 7.67 (d, 1H) 8.07 (s, 1H) 12.46 (br. s., 1H). LCMS:[M+H]⁺=273.1, Rt⁽³⁾=1.30 min.

6′-Methoxy-5′-methyl-4-oxo-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-3-carboxylic acid methyl ester

To a stirred suspension of sodium hydride (60%, 153 g, 6.36 mmol) indimethyl carbonate (3.82 mL, 45.4 mmol) at room temperature was slowlyadded 6′-methoxy-5′-methyl-2,3,5,6-tetrahydro-[1,3′]bipyridinyl-4-one (1g, 4.54 mmol). The reaction mixture was heated to reflux (90° C.) for 1h and then cooled to room temperature. The mixture was partitionedbetween CH₂Cl₂ and water and a solution of 1N HCl was added cautiously.The aqueous layer was separated and washed with an addition portion ofCH₂Cl₂. The combined organic extracts were washed with brine, dried oversodium sulfate, filtered and evaporated to give the crude product, whichwas purified by flash-chromatography on silica gel (heptane/ethylacetate3/1) to afford6′-methoxy-5′-methyl-4-oxo-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-3-carboxylicacid methyl ester (975 mg, yield 77%) as a white solid. ¹H-NMR (400 MHz,DMSO, 298K) (mixture of keto and enol tautomers observed) δ ppm 2.12 (s,6H) 2.36-2.69 (m, 4H) 3.26-3.96 (m, 20H) 7.34-7.77 (m, 4H) 11.84 (s,1H). LCMS: [M+H]⁺=279.1, Rt⁽³⁾=1.51 min (tautomer 1) and 1.70 min(tautomer 2).

6′-Methoxy-5′-methyl-2, 3, 5, 6-tetrahydro-[1, 3′]bipyridinyl-4-one

A slurry of iodide salt 1-benzyl-1-methyl-4-oxo-piperidinium (Ref:Tortolani, R.; Org. Lett., Vol. 1, No 8, 1999) (3.61 g, 10.86 mmol) inwater (10 mL) was added slowly to a refluxing solution of2-methoxy-5-amino-3-picolin (1 g, 7.24 mmol) and potassium carbonate(0.140 g, 1.013 mmol) in ethanol (20 mL). The reaction mixture washeated to reflux for an additional 3 h. The reaction mixture was cooledto rt and partitioned between CH₂Cl₂ and water. The organic layer wasseparated and washed with an addition portion of CH₂Cl₂. The combinedorganic layers were washed with brine, dried over sodium sulfate,filtered and concentrated to give the crude product which was purifiedby flash-chromatography on silica gel (heptane/ethylacetate 1/1) toafford 6′-methoxy-5′-methyl-2,3,5,6-tetrahydro-[1,3′]bipyridinyl-4-one(1.15 g, yield 72%) as a light yellow gum. ¹H-NMR (400 MHz, DMSO, 298K)δ ppm 2.12 (s, 3H) 2.42 (t, 4H) 3.46 (t, 4H) 3.80 (s, 3H) 7.40 (d, 1H)7.71 (d, 1H). LCMS: [M+H]⁺=221.1, Rt⁽³⁾=1.41 min.

Synthesis of Example 1—Method 1b (According to Scheme 1)

Step 3

To a mixture6-(6-methoxy-5-methyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(639 g, 1.87 mmol) in CH₂Cl₂ (5 mL) was added the acid chloridetetrahydro-2H-pyran-4-carbonyl chloride (306 g, 2.06 mmol) andtriethylamine (0.522 mL, 3.74 mmol) at rt. The reaction mixture wasstirred at rt for 10 min. The reaction mixture was concentrated undervacuum. Purification by preparative reverse phase Gilson HPLC andsubsequent neutralization of the combined fractions by extraction withCH₂Cl₂/1N NaOH, separation of the organic phase through a phaseseparation tube and evaporated gave{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone(432 mg, 51% yield) as a white powder. ¹H-NMR (400 MHz, DMSO-d6, 298K) δppm 1.50-1.65 (m, 4H) 2.10-2.32 (m, 5H) 2.62-2.78 (m, 1H) 2.85-2.95 (m,2H) 3.30-3.95 (m, 13H) 4.0-4.20 (m, 2H) 5.61-5.72 (m, 1H) 7.42 (br, 1H)7.68 (m, 1H) 8.60-8.61 (m, 1H). LCMS: [M+H]⁺=454.2, Rt⁽¹⁾=1.42 min.

6-(6-Methoxy-5-methyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

Step 2

(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (2.05 g, 4.63 mmol) was dissolved in TFA/CH₂Cl₂(1/2) and stirred at rt for 1 h. The reaction mixture was concentratedunder vacuum, the residue was diluted with CH₂Cl₂, the organic layerwashed with NaOH 1N then brine, dried over Na₂SO₄, filtered andevaporated to give6-(6-methoxy-5-methyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine.¹H NMR (400 MHz, CDCl₃, 298K) δ ppm 2.20-2.30 (m, 2H), 2.22 (s, 3H),3.00-3.06 (t, 2H), 3.09-3.18 (m, 1H), 3.22-3.37 (m, 3H), 3.45-3.50 (t,2H), 3.95 (s 3H), 4.10 (s, 2H), 4.20-4.65 (br. s 1H), 5.63-5.69 (m, 1H),7.21-7.252 (m, 1H), 7.70-7.74 (m, 1H), 8.60 (s, 1H). LCMS: [M+H]⁺=341.9,Rt⁽⁷⁾=0.61 min.

(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

Step 1

X-Phos (0.96 g, 2.01 mmol, 0.3 eq.), Pd₂(dba)₃ (0.615 g, 0.672 mmol, 0.1eq.), Cs₂CO₃ (4.38 g, 13.44 mmol, 2 eq.) were combined and flushed 10min with Argon. To this mixture, a solution of(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 7) (2.15 g, 6.72 mmol) in dioxane (6mL) and 5-bromo-2-methoxy-3-methylpyridine (1.76 g, 8.73 mmol) wereadded at rt and the reaction mixture was stirred at 120° C. for 2 h. Thereaction was cooled down to rt, the reaction mixture filtered overHyflo, AcOEt was added and the organic layer was washed with brine,dried over Na₂SO₄, filtered and concentrated under vacuum. The residuewas dissolved in dioxane (6 mL) and added to a glass vial containing5-bromo-2-methoxy-3-methylpyridine (1.76 g, 8.73 mmol), X-Phos (0.96 g,2.01 mmol), Pd₂(dba)₃ (0.615 g, 0.672 mmol), Cs₂CO₃ (4.38 g, 13.44mmol). The vial was capped and the reaction mixture was stirred at 120°C. for 2 h. The reaction was cooled down to rt, the reaction mixturefiltered over Hyflo, AcOEt was added and the organic layer was washedwith brine, dried over Na₂SO₄, filtered and concentrated under vacuum.Purification by flash chromatography on silica gel (CH₂Cl₂ then TBMEthen TBME/MeOH 99/1 to 90/10) gave(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester as a yellow foam (2.05 g, 69% yield). ¹H NMR (400MHz, DMSO-d6, 298K) δ ppm 1.35-1.44 (br. s., 9H) 2.07-2.23 (m, 2H), 2.14(s, 3H), 2.87-2.93 (m, 2H), 3.39-3.68 (m, 6H), 3.81 (s, 3H), 4.03-4.08(m, 2H), 5.56-5.63 (m, 1H), 7.41-7.46 (m, 1H), 7.67-7.73 (m, 1H), 8.60(s, 1H). LCMS: [M+H]⁺=342.2, Rt⁽²⁾=0.94 min.

Crystallization of Example 1 by Heating and Cooling in Acetonitrile

1 part of Example 1 (eg. 100 mg) was mixed with 5 parts of acetonitrile(0.5 mL for each 100 mg of compound) with stirring. A solution wasobtained by heating up to 40-60° C. The mixture was then allowed toslowly cooled down to RT. After further cooling overnight (5° C.),precipitation was observed. In case no precipitation is not observed,the volume of ethanol can be reduce using a nitrogen stream andrepeating the overnight cooling step. The mixture was centrifuged toremove the ethanol. The solid was dried under vacuum at 25° C. and 70mbar. A crystalline anhydrous form of Example 1 with a MP of 131° C. wasobtained. This crystalline form was also observed under other methodsand/or solvents, such as heating and cooling in ethanol, acetone, ethylacetate, isopropanol, by slurry in heptane, or by antisolvent additionin THF or 3-methyl-1-butanol using heptane as antisolvent. These resultsshow the reproducibility and scalability of the crystalline form as wellas suggests that the same form can be prepared under differentexperimental conditions than the ones described above.

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 1 anhydrous form (Method X2):

2-Theta in deg Intensity in % 7.5 56 10.9 12.5 11.7 25.1 14.3 23.8 15.1100 15.8 40.9 16.7 22.1 17.7 65.1 18.9 28.9 20.5 24.7 21.8 26 22.5 28.323.3 31.3 24.2 76.1 24.6 51.8 25.0 41.3 25.6 20.4 26.2 20.8 27.0 14.228.0 17.5 29.1 16.1 32.8 14 34.6 11.4

Crystallization of Trihydrate Form of Example 1 by Slurry in Water

Slurry of Example 1 in water e.g., 1 part of Example 1 in 10 parts ofwater, at RT produced a trihydrate form of Example 1. The crystals wereseparated by centrifugation and dried at room environment.

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 1 trihydrate form (Method X2):

2-Theta in deg Intensity in % 6.6 24.3 8.9 7.9 13.3 100 14.5 18.3 15.012.6 16.5 12.4 17.5 15.7 17.7 17.2 18.2 9.8 20.0 10.7 21.6 11.7 22.620.3 23.8 11.4 24.4 15.2 26.7 26.5 27.5 18.7 27.8 16.6 29.2 9.8 33.3 933.9 7.6 35.7 8.2 38.8 7

Preparation of Citrate Salt of Example 1

0.5 g of Example 1 (assay 91.8%) were dissolved in 5 mL ofmethylethylketone and 0.25 mL of water and heated at 60° C. 213 mg ofcitric acid were added at 50° C. and the mixture was allowed to cooldown to RT within 30 min. Crystallization occurs at 45° C. The mixturewas stirred for 16 h at RT. The crystals were collected by filtration.The filter cake was washed 3 times with 1 mL of methylethylketone andafterwards dried for 16 h at 50° C. and ca. 10 mbar vacuum. Elementaryanalysis of the citrate salt showed a 1:1 (monohydrate) form.

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 1 citrate salt (Method X1):

2-Theta in deg Intensity in % 5.7 62 11.5 100 12.1 4 14.3 4 15.4 12 17.221 17.9 31 19.3 25 20.2 37 20.7 8 21.9 5 23.3 11 23.9 36 25.5 28 27.0 527.7 6 29.8 8 30.3 7

Preparation of Fumarate Salt of Example 1

0.5 g Example 1 (assay 91.8%) were dissolved in 15 mL of acetonitrileand 0.2 mL of water and heated at 76° C. 129 mg of fumaric acid wereadded at 60° C. The solution was allowed to cool down to RT within 30min. The salt precipitated and the suspension was stirred for 16 h atRT. The crystals were collected by filtration. The filter cake waswashed 3 times with 1 mL of acetonitrile and afterwards dried for 16 hat 50° C. and ca. 10 mbar vacuum. Elementary analysis of the fumaratesalt showed a 1:1 (monohydrate) form.

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 1 fumarate salt (Method X1):

2-Theta in deg Intensity in % 6.0 100 6.5 12 9.8 8 12.3 10 13.1 14 15.622 17.7 16 19.1 21 19.7 27 23.9 40 24.7 6 24.9 10 25.2 5 26.4 11 27.0 4

Preparation of Napadisylate Salt of Example 1

0.5 g Example 1 (assay 91.8%) were dissolved in 5 mL of ethanol absoluteand 0.25 mL of water at 60° C. 250 mg of naphthalendisulfonic acid wereadded at 50° C. and the mixture was allowed to cool down to RT within 30min. Crystallization occurs at 40° C. The mixture was stirred for 16 hat RT. The crystals were collected by filtration. The filter cake waswashed 3 times with 1 mL of ethanol and afterwards dried for 16 h at 50°C. and ca. 10 mbar vacuum. Elementary analysis of the napadisylate saltshowed a 2:1 (monohydrate) form.

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 1 napadisylate salt (Method X1):

2-Theta in deg Intensity in % 4.3 100 8.5 3 9.4 6 12.2 12 12.9 12 13.537 15.0 26 15.6 12 16.0 11 17.7 28 18.9 23 19.3 11 20.0 11 20.8 3 21.2 522.0 9 23.0 41 24.5 39 26.5 20

Examples 2-9 were prepared using procedures analogous to those used inExample 1 (method 1b) using appropriate starting materials.

Example 2

Rt⁽¹⁾ (min.) 471.3 MS: [M + H]+ 1.21

Name:{(S)-3-[6-(2,4-Dimethoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase method C

Prepared using process step 3 of method 1b from intermediate 8 andtetrahydro-pyran-4-carbonyl chloride

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 1.50-1.86 (m, 4H) 2.20-2.45(m, 2H) 2.70-2.87 (m, 1H) 2.96-2.99 (m, 2H) 3.35-4.14 (m, 18H) 5.69-5.85(m, 1H) 7.96 (m, 1H) 8.58 (m, 1H)

Example 3

Rt⁽¹⁾ (min.) 1.42 MS: [M + H]+ 465.2

Name:2-Methoxy-5-{4-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Purification method: Reverse phase method A

Prepared using process steps 2-3 of method 1b from intermediate 11 andtetrahydro-pyran-4-carbonyl chloride

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.50-1.64 (m, 4H) 2.10-2.31 (m,2H) 2.62-2.77 (m, 1H) 2.87-2.95 (m, 2H) 3.29-3.96 (m, 13H) 4.08-4.21 (m,2H) 5.58-5.73 (m, 1H) 8.06-8.09 (m, 1H) 8.23-8.27 (m, 1H) 8.60-8.64 (m,1H)

Example 4

Rt⁽¹⁾ (min.) 1.27 MS: [M + H]+ 414.2

Name:1-{(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-propan-1-one

Purification method: Reverse phase method A

Prepared using process steps 2-3 of method 1b from intermediate 10 andpropionyl chloride

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 1.10-1.20 (m, 3H) 2.19-2.49 (m,4H) 3.02-3.08 (m, 2H) 3.45-3.52 (m, 2H) 3.56-3.68 (m, 2H) 3.72-3.90 (m,2H) 3.91 (s, 3H) 3.99 (s, 3H) 4.07-4.12 (m, 2H) 5.75-5.78 (m, 1H)6.89-7.01 (m, 1H) 7.44-7.46 (m, 1H) 8.60-8.62 (m, 1H)

Example 5

Rt⁽¹⁾ (min.) 1.25 MS: [M + H]+ 470.2

Name:{(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using process steps 2-3 of method 1b from intermediate 10 andtetrahydro-pyran-4-carbonyl chloride

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 1.56-1.68 (m, 2H) 1.88-2.00 (m,2H) 2.20-2.38 (m, 2H) 2.53-2.70 (m, 1H) 3.05-3.10 (m, 2H) 3.39-3.54 (m,4H) 3.59-3.82 (m, 4H) 3.91 (s, 3H) 3.99 (s, 3H) 4.01-4.10 (m, 4H)5.62-5.78 (m, 1H) 6.89-6.90 (m, 1H) 7.40-7.43 (m, 1H) 8.60-8.65 (m, 1H)

Example 6

Rt⁽¹⁾ (min.) 1.10 MS: [M + H]+ 450.2

Name:2-Amino-5-{4-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Prepared using process step 3 of method 1b from intermediate 9 andtetrahydro-pyran-4-carbonyl chloride

Purification method: 1—Normal phase chromatography CH₂Cl₂/MeOH assolvent

-   -   2—Reverse phase method A

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 1.45-1.75 (m, 2H) 1.86-2.02 (m,2H) 2.20-2.40 (m, 2H) 2.50-2.75 (m, 1H) 3.02-3.09 (m, 2H) 3.38-4.20 (m,12H) 4.96 (s, 1H) 5.70-5.78 (m, 1H) 7.39 (m, 1H) 8.13-8.14 (m, 1H)8.62-8.64 (m, 1H)

Example 7

Rt⁽¹⁾ (min.) 1.41 MS: [M + H]+ 458.1

Name:{(S)-3-[6-(5-Fluoro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Biotage 11 g KP—NH cartridge eluting withHeptane/EtOAc 100/0 to 0/100

Prepared using process steps 2-3 of method 1b from intermediate 12 andtetrahydro-pyran-4-carbonyl chloride

¹H NMR (400 MHz, CDCl₃, 298K) δ ppm 1.56-1.74 (m, 2H) 1.87-2.02 (m, 2H)2.19-2.42 (m, 2H) 2.51-2.74 (m, 1H) 3.01-3.09 (m, 2H) 3.39-4.20 (m, 15H)5.70-5.79 (m, 1H) 7.13-7.20 (m, 1H) 7.63-7.69 (m, 1H) 8.59-8.66 (m, 1H)

Example 8

Rt⁽¹⁾ (min.) 1.35 MS: [M + H]+ 425.1

Name:2-Methoxy-5-{4-[(S)-1-(2-methoxy-acetyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Purification method: Reverse phase method A

Prepared using process steps 2-3 of method 1b from intermediate 11 andmethoxy acetyl chloride

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.11-2.32 (m, 2H) 2.88-2.95 (m,2H) 3.26-3.32 (m, 3H) 3.46-3.84 (m, 6H) 3.91-3.95 (m, 3H) 3.98-4.08 (m,2H) 4.13-4.19 (m, 2H) 5.59-5.71 (m, 1H) 8.07-8.10 (m, 1H) 8.25-8.28 (m,1H) 8.61-8.62 (m, 1H)

Example 9

Rt⁽⁸⁾ (min.) 3.79 MS: [M + H]+ 449.1

Name:5-[4-((S)-1-Cyclopentanecarbonyl-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]-2-methoxy-nicotinonitrile

Purification method: Reverse phase method A

Prepared using process steps 2-3 of method 1b from intermediate 11 andcyclopentanecarbonyl chloride

Example 10:(2,4-Dimethyl-oxazol-5-yl)-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Step 1

A mixture of 2,4-dimethyl-oxazole-5-carboxylic acid (36.4 g, 0.258mmol), HTBU (98 g, 0.258 mmol), DIPEA (86μl, 0.49 mmol) in DMF (5 mL)was stirred at rt for 20 min. then a solution of6-(6-methoxy-5-methyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(prepared in example 1, method Ib, step 2) (80 g, 0.23 mmol) and DIPEA(86μl, 0.49 mmol) in DMF (0.4 mL) was added. The reaction mixture wasstirred 30 min at rt. The reaction mixture was directly purified bypreparative reverse phase Gilson HPLC and subsequent neutralization ofthe combined fractions over PL-HCO₃ MP gave(2,4-dimethyl-oxazol-5-yl)-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone(91 mg, 84% yield) as a white lyophilized powder. ¹H-NMR (400 MHz,methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.27-2.52 (m, 8H) 2.95-3.03 (m,2H) 3.44-3.55 (m, 2H) 3.70-4.26 (m, 9H) 5.76-5.92 (m, 1H) 7.40 (br. s.,1H) 7.64 (br. s., 1H) 8.55-8.62 (m, 1H), LCMS: [M+H]+=465.2, Rt⁽¹⁾=1.51min.

Examples 11-49 and 51-53 were prepared using procedures analogous tothose used in Example 10, step 1 using appropriate starting materials.

Example 11

Rt⁽¹⁾ (min.) 1.57 MS: [M + H]+ 436.2

Name:Furan-3-yl-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using furan-3-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.30-2.45 (m, 2H)2.93-3.05 (m, 2H) 3.45-3.54 (m, 2H) 3.72-4.21 (m, 9H) 5.79-5.86 (m, 1H)6.78-6.82 (m, 1H) 7.37-7.44 (m, 1H) 7.56-7.61 (m, 1H) 7.61-7.69 (m, 1H)8.01-8.12 (m, 1H) 8.54-8.62 (m, 1H)

Example 12

Rt⁽¹⁾ (min.) 1.36 MS: [M + H]+ 437.2

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-5-yl-methanone

Purification method: Reverse phase method A

Prepared using oxazole-5-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.30-2.39 (m, 1H)2.41-2.50 (m, 1H) 2.95-3.03 (m, 2H) 3.45-3.52 (m, 2H) 3.76-4.32 (m, 9H)5.79-5.94 (m, 1H) 7.40 (br. s., 1H) 7.62-7.66 (m, 1H) 7.75-7.82 (m, 1H)8.34-8.40 (m, 1H) 8.56-8.61 (m, 1H)

Example 13

Rt⁽¹⁾ (min.) 1.13 MS: [M + H]+ 450.2

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-3H-imidazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 3-methyl-3H-imidazole-4-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.30-2.45 (m, 2H)2.93-3.05 (m, 2H) 3.43-3.55 (m, 2H) 3.74-4.24 (m, 12H) 5.82 (br. s., 1H)7.35-7.56 (m, 2H) 7.66 (m, 1H) 7.76 (br. s., 1H) 8.55-8.60 (m, 1H)

Example 14

Rt⁽¹⁾ (min.) 1.49 MS: [M + H]+ 451.2

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2-methyl-oxazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 2-methyl-oxazole-4-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.27-2.43 (m, 2H)2.43-2.50 (m, 3H) 2.95-3.02 (m, 2H) 3.45-3.53 (m, 2H) 3.72-4.33 (m, 9H)5.78-5.89 (m, 1H) 7.37-7.43 (m, 1H) 7.61-7.67 (m, 1H) 8.25-8.31 (m, 1H)8.57-8.60 (m, 1H)

Example 15

Rt⁽¹⁾ (min.) 1.53 MS: [M + H]+ 454.2

Name:(3-Methoxy-cyclobutyl)-{(S)-3-[6-(6-methoxy-1-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using 3-methoxy-cyclobutanecarboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 1.99-2.55 (m, 9H) 2.78-2.95(m, 1H), 2.95-3.02 (m, 2H) 3.20-3.23 (m, 3H) 3.47-3.52 (m, 2H) 3.52-4.10(m, 10H) 5.73-5.81 (m, 1H) 7.38-7.42 (m, 1) 7.63-7.67 (m, 1H) 8.57 (s,1H)

Example 16

Rt⁽¹⁾ (min.) 1.41 MS: [M + H]+ 437.2

Name:({(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanone

Purification method: Reverse phase method A

Prepared using oxazole-4-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.29-2.37 (m, 1H)2.37-2.44 (m, 1H) 2.94-3.03 (m, 2H) 3.45-3.53 (m, 2H) 3.75-4.38 (m, 9H)5.79-5.89 (m, 1H) 7.38-7.42 (m, 1H) 7.62-7.66 (m, 1H) 8.19-8.26 (m, 1H)8.44-8.48 (m, 1H) 8.56-8.61 (m, 1H)

Example 17

Rt⁽¹⁾ (min.) 1.35 MS: [M + H]+ 495.2

Name:1-(4-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanone

Purification method: Reverse phase method A

Prepared using 1-acetyl-piperidine-4-carboxylic acid

¹H NMR ((400 MHz, methanol-d4, 298K) δ ppm 1.49-1.89 (m, 4H) 2.06-2.13(m, 3H) 2.18 (s, 3H) 2.23-2.43 (m, 2H) 2.61-2.93 (m, 2H) 2.95-3.04 (m,2H) 3.15-3.25 (m, 1H) 3.42-3.53 (m, 2H) 3.55-4.12 (m, 10H) 4.46-4.59 (m,1H) 5.74-5.86 (m, 1H) 7.38-7.45 (m, 1H) 7.62-7.67 (m, 1H) 8.56-8.61 (m,1H)

Example 18

Rt⁽¹⁾ (min.) 1.47 MS: [M + H]+ 451.2

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(4-methyl-oxazol-5-yl)-methanone

Purification method: Reverse phase method A

Prepared using 4-methyl-oxazole-5-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.29-2.47 (m, 5H)2.95-3.3.03 (m, 2H) 3.45-3.52 (m, 2H) 3.73-4.30 (m, 9H) 5.79-5.90 (m,1H) 7.41 (m, 1H) 7.65 (br. s., 1H) 8.19-8.24 (m, 1H) 8.55-8.61 (m, 1H)

Example 19

Rt⁽¹⁾ (min.) 1.25 MS: [M + H]+ 463.1

Name:5-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-1H-pyridin-2-one

Purification method: Reverse phase method A

Prepared using 6-oxo-1,6-dihydro-pyridine-3-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.19 (s, 3H) 2.30-2.40 (m, 2H)2.95-3.05 (m, 2H) 3.45-3.55 (m, 2H) 3.74-4.22 (m, 9H) 5.73-5.85 (m, 1H)6.50-6.56 (m, 1H) 7.39-7.45 (m, 1H) 7.60-7.70 (m, 1H) 7.78-7.90 (m, 2H)8.50-8.60 (m, 1H)

Example 20

Rt⁽¹⁾ (min.) 1.22 MS: [M + H]+ 450.2

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 1-methyl-1H-imidazole-4-carboxylic acid

¹H NMR (400 MHz, methanol-d4, 298K) δ ppm 2.17 (s, 3H) 2.28-2.41 (m, 2H)2.94-3.02 (m, 2H) 3.45-3.52 (m, 2H) 3.73-4.35 (m, 12H) 5.80-5.85 (m, 1H)7.38-7.43 (m, 1H) 7.60-7.69 (m, 3H) 8.55-8.61 (m, 1H)

Example 21

Rt⁽¹⁾ (min.) 1.23 MS: [M + H]+ 453.1

Name:{(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanone

Purification method: Normal phase chromatography with EtOAc/MeOH assolvent

Prepared using intermediate 10 and method 1b of process step 2 ofexample 1 followed by process step 1 of example 10 usingoxazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.10-2.37 (m, 2H) 2.81-2.99 (m,2H) 3.46-4.27 (m, 14H) 5.58-5.77 (m, 1H) 7.08-7.20 (m, 1H) 7.30-7.42 (m,1H) 8.43-8.54 (m, 1H) 8.55-8.69 (m, 2H)

Example 22

Rt⁽¹⁾ (min.) 1.18 MS: [M + H]+ 453.1

Name:{(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-5-yl-methanone

Purification method: Reverse phase method A

Prepared using intermediate 10 and process step 2, method 1b of example1 followed by process step 1 of example 10 using oxazole-5-carboxylicacid

¹H NMR (400 MHz, DMSO-d6, 373K) δ ppm 2.22-2.42 (m, 2H) 2.80-3.00 (m,2H) 3.50-4.30 (m, 14H) 5.63-5.83 (m, 1H) 7.06-7.09 (m, 1H) 7.38-7.40 (m,1H) 7.69 (s, 1H) 8.40 (s, 1H) 8.57 (s, 1H)

Example 23

Rt⁽¹⁾ (min.) 1.3 MS: [M + H]+ 467.2

Name:{(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2-methyl-oxazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 10 and process step 2, method 1b of example1 followed by process step 1 of example 10 using2-methyl-oxazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.11-2.36 (m, 2H) 2.40-2.44 (m,3H) 2.81-2.97 (m, 2H) 3.40-4.28 (m, 14H) 5.62-5.78 (m, 1H) 7.11-7.21 (m,1H) 7.29-7.41 (m, 1H) 8.42-8.52 (m, 1H) 8.59-8.67 (m, 1H)

Example 24

Rt⁽¹⁾ (min.) 1.38 MS: [M + H]+ 498.3

Name: {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2,2-dimethyl-tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 10 and process step 2, method 1b of example1 followed by process step 1 of example 10 using2,2-dimethyl-tetrahydro-pyran-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 0.97-1.24 (m, 6H) 1.28-1.58 (m,4H) 2.08-2.34 (m, 2H) 2.72-2.97 (m, 3H) 3.43-4.12 (m, 16H) 5.55-5.76 (m,1H) 7.14-7.20 (m, 1H) 7.31-7.37 (m, 1H) 8.59-8.64 (m, 1H)

Example 25

Rt⁽¹⁾ (min.) 1.32 MS: [M + H]+ 481.2

Name: {(S)-3-[6-(5,6-Dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2,4-dimethyl-oxazol-5-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 10 and process step 2, method 1b of example1 followed by process step 1 of example 10 using2,2-dimethyl-tetrahydro-pyran-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.11-2.48 (m, 8H) 2.83-2.98 (m,2H) 3.43-4.18 (m, 14H) 5.56-5.89 (m, 1H) 7.12-7.20 (m, 1H) 7.32-7.40 (m,1H) 8.57-8.67 (m, 1H)

Example 26

Rt⁽¹⁾ (min.) 1.57 MS: [M + H]+ 504.3

Name:(4,4-Difluoro-cyclohexyl)-{(S)-3-[6-(5,6-dimethoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using intermediate 10 and process step 2, method 1b of example1 followed by process step 1 of example 10 using4,4-difluoro-cyclohexanecarboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.49-2.35 (m, 10H) 2.61-2.67 (m,1H) 2.84-2.99 (m, 2H) 3.42-3.83 (m, 12H) 4.00-4.19 (m, 2H) 5.57-5.78 (m,1H) 7.11-7.25 (m, 1H) 7.29-7.43 (m, 1H) 8.52-8.68 (m, 1H)

Example 27

Rt⁽¹⁾ (min.) 1.46 MS: [M + H]+ 479.2

Name:2-Methoxy-5-{4-[(S)-1-(2-tetrahydro-pyran-4-yl-acetyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Purification method: Reverse phase method A

Prepared using intermediate 11 and process step 2, method 1b of example1 followed by process step 1 of example 10 using(tetrahydro-pyran-4-yl)-acetic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.11-1.28 (m, 2H) 1.49-1.64 (m,2H) 1.87-1.99 (m, 1H) 2.07-2.29 (m, 4H) 2.86-2.95 (m, 2H) 3.19-3.30 (m,2H) 3.42-3.88 (m, 8H) 3.90-3.96 (m, 3H) 4.09-4.19 (m, 2H) 5.57-5.70 (m,1H) 8.07-8.11 (m, 1H) 8.22-8.28 (m, 1H) 8.58-8.65 (m, 1H)

Example 28

Rt⁽¹⁾ (min.) 1.51 MS: [M + H]+ 476.2

Name:5-{4-[(S)-1-(2,4-Dimethyl-oxazole-5-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-2-methoxy-nicotinonitrile

Purification method: Reverse phase method A

Prepared using intermediate 11 and process step 2, method 1b of example1 followed by process step 1 of example 10 using2,4-dimethyl-oxazole-5-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.16-2.46 (m, 5H) 2.30 (s, 3H)2.85-2.96 (m, 2H) 3.50-4.20 (m, 8H) 3.92 (s, 3H) 5.64-5.80 (m, 1H)8.04-8.12 (m, 1H) 8.22-8.30 (m, 1H) 8.62 (s, 1H)

Example 29

Rt⁽¹⁾ (min.) 1.58 MS: [M + H]+ 493.2

Name:5-{4-[(S)-1-(2,2-Dimethyl-tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-2-methoxy-nicotinonitrile

Purification method: Reverse phase method A

Prepared using intermediate 11 and process step 2, method 1b of example1 followed by process step 1 of example 10 using2,2-dimethyl-tetrahydro-pyran-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.00-1.24 (m, 6H) 1.28-1.73 (m,4H) 2.10-2.34 (m, 2H) 2.62-2.97 (m, 3H) 3.43-3.84 (m, 8H) 3.94 (s, 3H)4.09-4.20 (m, 2H) 5.58-5.75 (m, 1H) 8.05-8.11 (m, 1H) 8.20-8.29 (m, 1H)8.59-8.65 (m, 1H)

Example 30

Rt⁽¹⁾ (min.) 1.55 MS: [M + H]+ 451.1

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(5-methyl-oxazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 5-methyl-oxazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.10-2.32 (m, 2H) 2.13 (s, 3H)2.52.2.54 (m, 3H) 2.85-2.93 (m, 2H) 3.42-3.50 (m, 2H) 3.61-4.22 (m, 6H)3.81 (s, 3H) 5.64-5.72 (m, 1H) 7.41-7.45 (m, 1H) 7.67-7.71 (m, 1H)8.27-8.33 (m, 1H) 8.59-8.64 (m, 1H)

Example 31

Rt⁽¹⁾ (min.) 1.53 MS: [M + H]+ 451.1

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(5-methyl-isoxazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 5-methyl-isoxazole-4-carboxylic acid ¹H NMR (400 MHz,DMSO-d6, 298K) 6 ppm 2.10-2.31 (m, 2H) 2.14 (s, 3H) 2.54-2.59 (m, 3H)2.83-2.97 (m, 2H) 3.41-3.53 (m, 2H) 3.59-4.15 (m, 6H) 3.81 (s, 3H)5.65-5.73 (m, 1H) 7.40-7.48 (m, 1H) 7.67-7.74 (m, 1H) 8.56-8.66 (m, 1H)8.83-8.95 (m, 1H)

Example 32

Rt⁽¹⁾ (min.) 1.53 MS: [M + H]+ 451.1

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-isoxazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 3-methyl-isoxazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.08-2.37 (m, 8H) 2.82-2.95 (m,2H) 3.40-3.53 (m, 2H) 3.55-4.16 (m, 9H) 5.65-5.75 (m, 1H) 7.41-7.48 (m,1H) 7.68-7.73 (m, 1H) 8.57-8.65 (m, 1H) 9.28-9.40 (m, 1H)

Example 33

Rt⁽¹⁾ (min.) 1.59 MS: [M + H]+ 437.2

Name:Isoxazol-3-yl-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using isoxazole-3-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.13 (m, 3H) 2.21-2.31 (m, 2H)2.86-2.94 (m, 2H) 3.43-3.50 (m, 2H) 3.66-4.15 (m, 9H) 5.67-5.73 (m, 1H)6.84-6.91 (m, 1H) 7.42-7.46 (m, 1H) 7.67-7.74 (m, 1H) 8.57-8.64 (m, 1H)9.05-9.13 (m, 1H)

Example 34

Rt⁽¹⁾ (min.) 1.5 MS: [M + H]+ 437.2

Name:Isoxazol-5-yl-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using isoxazole-5-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.08-2.16 (m, 3H) 2.19-2.36 (m,2H) 2.85-2.95 (m, 2H) 3.42-3.49 (m, 2H) 3.66-4.23 (m, 9H) 5.66-5.78 (m,1H) 7.06-7.13 (m, 1H) 7.41-7.46 (m, 1H) 7.68-7.74 (m, 1H) 8.59-8.64 (m,1H) 8.73-8.79 (m, 1H)

Example 35

Rt⁽¹⁾ (min.) 1.5 MS: [M + H]+ 464.1

Name:2-Methoxy-5-{4-[(S)-1-(thiazole-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Purification method: Reverse phase method A

Prepared using intermediate 11 and process step 2, method 1b of example1 followed by process step 1 of example 10 using thiazole-4-carboxylicacid

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 2.30-2.37 (m, 2H) 3.07-3.12 (m,2H) 3.46-3.53 (m, 2H) 3.81-4.43 (m, 9H) 5.80-5.85 (m, 1H) 7.55-7.59 (m,1H) 8.09-8.13 (m, 1H) 8.18-8.23 (m, 1H) 8.63-8.69 (m, 1H) 8.75-8.85 (m,1H)

Example 36

Rt⁽¹⁾ (min.) 1.35 MS: [M + H]+ 461.2

Name:2-Methoxy-5-{4-[(S)-1-(1-methyl-1H-pyrazole-4-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Purification method: Reverse phase method A

Prepared using intermediate 11 and process step 2, method 1b of example1 followed by process step 1 of example 10 using1-methyl-1H-imidazole-4-carboxylic acid

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 2.24-2.47 (m, 2H) 3.03-3.17 (m,2H) 3.45-3.58 (m, 2H) 3.87-4.20 (m, 12H) 5.75-5.85 (m, 1H) 7.54-7.60 (m,1H) 7.73-7.90 (m, 2H) 8.09-8.14 (m, 1H) 8.61-8.68 (m, 1H)

Example 37

Rt⁽¹⁾ (min.) 1.47 MS: [M + H]+ 461.2

Name:2-Methoxy-5-{4-[(S)-1-(1-methyl-1H-pyrazole-3-carbonyl)-pyrrolidin-3-yloxy]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Purification method: Reverse phase method A

Prepared using intermediate 11 and process step 2, method 1b of example1 followed by process step 1 of example 10 using1-methyl-1H-pyrazole-3-carboxylic acid

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 2.24-2.41 (m, 2H) 3.07-3.14 (m,2H) 3.44-3.58 (m, 2H) 3.74-4.44 (m, 12H) 5.77-5.86 (m, 1H) 6.78-6.84 (m,1H) 7.33-7.39 (m, 1H) 7.54-7.59 (m, 1H) 8.08-8.14 (m, 1H) 8.63-8.70 (m,1H)

Example 38

Rt⁽¹⁾ (min.) 1.58 MS: [M + H]+ 482.3

Name:(2,2-Dimethyl-tetrahydro-pyran-4yl)-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using 2,2-dimethyl-tetrahydro-pyran-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.02-1.21 (m, 6H) 1.27-1.71 (m,4H) 2.08-2.32 (m, 5H) 2.67-2.94 (m, 3H) 3.41-4.08 (m, 13H) 5.60-5.73 (m,1H) 7.41-7.46 (m, 1H) 7.65-7.72 (m, 1H) 8.58-8.65 (m, 1H)

Example 39

Rt⁽¹⁾ (min.) 1.60 MS: [M + H]+ 556.1

Name:(1,1-Dioxo-hexahydro-1lambda*6*-thiopyran-4-yl)-{(S)-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using1,1-dioxo-hexahydro-1lambda*6*-thiopyran-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.90-2.37 (m, 6H) 2.72-3.27 (m,7H) 3.43-3.81 (m, 6H) 3.89-3.97 (m, 3H) 4.13-4.20 (m, 2H) 5.61-5.75 (m,1H) 7.80-7.86 (m, 1H) 8.15-8.22 (m, 1H) 8.60-8.65 (m, 1H)

Example 40

Rt⁽¹⁾ (min.) 1.77 MS: [M + H]+ 519.2

Name:(2,4-Dimethyl-oxazol-5-yl)-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using2,4-dimethyl-oxazole-5-carboxylic acid ¹H NMR (400 MHz, DMSO-d6, 298K) δppm 2.13-2.45 (m, 8H) 2.89-2.96 (m, 2H) 3.54-4.21 (m, 11H) 5.64-5.79 (m,1H) 7.81-7.85 (m, 1H) 8.218-8.22 (m., 1H) 8.61-8.65 (m, 1H)

Example 41

Rt⁽¹⁾ (min.) 1.69 MS: [M + H]+ 507.1

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-thiazol-5-yl-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using thiazole-5-carboxylicacid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.15-2.42 (m, 2H) 2.88-2.97 (m,2H) 3.53-3.61 (m, 2H) 3.67-4.11 (m, 7H) 4.15-4.24 (m, 2H) 5.67-5.79 (m,1H) 7.81-7.88 (m, 1H) 8.18-8.23 (m, 1H) 8.35-8.45 (m, 1H) 8.60-8.66 (m,1H) 9.22-9.29 (m, 1H)

Example 42

Rt⁽¹⁾ (min.) 1.74 MS: [M + H]+ 504.2

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(2-methyl-2H-pyrazol-3-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using2-methyl-2H-pyrazole-3-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.15-2.35 (m, 2H) 2.88-2.97 (m,2H) 3.51-4.13 (m, 12H) 4.13-4.25 (m, 2H) 5.63-5.74 (m, 1H) 6.63-6.74 (m,1H) 7.43-7.52 (m, 1H) 7.81-7.89 (m, 1H) 8.17-8.25 (m, 1H) 8.57-8.67 (m,1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 43

1.47 507.2

Name:4-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-pyrrolidin-2-one

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using5-oxo-pyrrolidine-3-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.10-2.43 (m, 4H) 2.89-2.96 (m,2H) 3.35-3.79 (m, 9H) 3.90-3.94 (m, 3H) 4.15-4.20 (m, 2H) 5.60-5.73 (m,1H) 7.53-7.62 (m, 1H) 7.81-7.87 (m, 1H) 8.17-8.22 (m, 1H) 8.60-8.64 (m,1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 44

1.63 501.2

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-pyridin-3-yl-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using nicotinic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.11-2.37 (m, 2H) 2.87-2.99 (m,2H) 3.51-4.13 (m, 9H) 4.13-4.29 (m, 2H) 5.60-5.75 (m, 1H) 7.43-7.53 (m,1H) 7.81-8.04 (m, 2H) 8.17-8.28 (m, 1H) 8.53-8.82 (m, 3H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 45

1.36 490.2

Name:(1H-Imidazol-4-yl)-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using1H-imidazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.12-2.35 (m, 2H) 2.87-2.95 (m,2H) 3.60-4.31 (m, 11H) 5.63-5.76 (m, 1H) 7.57-7.65 (m, 1H) 7.70-7.78 (m,1H) 7.80-7.85 (m, 1H) 8.16-8.21 (m, 1H) 8.61-8.65 (m, 1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 46

1.47 507.2

Name:5-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-pyrrolidin-2-one

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using5-oxo-pyrrolidine-2-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.78-2.40 (m, 6H) 2.89-2.97 (m,2H) 3.43-3.86 (m, 6H) 3.90-3.94 (m, 3H) 4.15-4.20 (m, 2H) 4.30-4.45 (m,1H) 5.60-5.75 (m, 1H) 7.70-7.89 (m, 2H) 8.16-8.22 (m, 1H) 8.61-8.63 (m,1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 47

1.61 501.2

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-pyridin-4-yl-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using isonicotinic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.12-2.36 (m, 2H) 2.86-2.99 (m,2H) 3.46-3.81 (m, 5H) 3.84-4.13 (m, 4H) 4.14-4.28 (m, 2H) 5.59-5.74 (m,1H) 7.44-7.56 (m, 2H) 7.82-7.91 (m, 1H) 8.18-8.27 (m, 1H) 8.52-8.72 (m,3H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 48

1.62 518.2

Name:(1,3-Dimethyl-1H-pyrazol-4-yl)-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using1,3,-dimethyl-1H-pyrazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.13-2.35 (m, 5H) 2.89-2.97 (m,2H) 3.53-4.06 (m, 12H) 4.14-4.22 (m, 2H) 5.64-5.72 (m, 1H) 7.80-7.88 (m,1H) 7.99-8.11 (m, 1H) 8.16-8.22 (m, 1H) 8.58-8.66 (m, 1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 49

1.51 490.1

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1H-pyrazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using1H-pyrazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.12-2.39 (m, 2H) 2.87-2.97 (m,2H) 3.43-4.11 (m, 9H) 4.12-4.22 (m, 2H) 5.63-5.79 (m, 1H) 7.78-7.94 (m,2H) 8.10-8.25 (m, 3H) 8.59-8.68 (m, 1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 51

1.72 506.2

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(5-methyl-[1,3,4]oxadiazol-2-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using5-methyl-[1,3,4]oxadiazole-2-carboxylic acid

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 52

1.71 502.1

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-pyrazin-2-yl-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using pyrazine-2-carboxylicacid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.15-2.36 (m, 2H) 2.87-2.97 (m,2H) 3.52-3.64 (m, 2H) 3.66-4.11 (m, 7H) 4.12-4.24 (m, 2H) 5.67-5.75 (m,1H) 7.81-7.87 (m, 1H) 8.17-8.25 (m, 1H) 8.56-8.80 (m, 3H) 8.97-9.02 (m,1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 53

1.45 504.2

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 13 and process step 2, method 1b of example1 followed by process step 1 of example 10 using1-methyl-1H-imidazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.07-2.36 (m, 2H) 2.82-3.05 (m,2H) 3.20-4.43 (m, 14H) 5.60-5.74 (m, 1H) 7.59-7.72 (m, 2H) 7.78-7.87 (m,1H) 8.14-8.21 (m, 1H) 8.59-8.66 (m, 1H)

Example 54:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-pyrazol-4-yl)-methanone

To a mixture of6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(prepared using step 1, example 91 from intermediate 13) (44 g, 0.11mmol), 1-methyl-1H-pyrazole-4-carboxylic acid (15 g, 0.12 mmol),benztriazol-1-ol (19 g, 0.12 mmol) in CH₂Cl₂ (1.0 mL) was added EDC (34g, 0.18 mmol) and the resulting mixture was stirred at rt for 18 h.Partitioned between CH₂Cl₂ (10 mL) and sat. NaHCO₃(aq) (2.0 mL) and theorganic layer separated by filtering through a phase separation tube.Concentrated in vacuo and purified by flash chromatography throughBiotage® amino silica gel eluting with cyclohexane/EtOAc, 100/0 to 0/100to give{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(1-methyl-1H-pyrazol-4-yl)-methanoneas a white lyophilized powder (44 mg, 75% yield). 1H NMR (400 MHz,CDCl₃-d, 298K) δ ppm 2.26-2.45 (m, 2H) 3.04-3.10 (m, 2H) 3.49-3.57 (m,2H) 3.89-4.00 (m, 7H) 4.01 (s, 3H) 4.10-4.18 (m, 2H) 5.78-5.83 (m, 1H)7.60-7.62 (m, 1H) 7.76-7.89 (m, 2H) 8.04-8.07 (m, 1H) 8.61-8.66 (m, 1H)MS: [M+H]⁺=504.2, Rt⁽³⁾=1.59 min.

Example 55 was prepared using procedures analogous to those used inexample 54 using appropriate starting materials.

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 55

1.77 507.2

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-thiazol-4-yl-methanone

Purification method: Flash chromatography on Biotage amino silica geleluting with cyclohexane/EtOAc 100/0 to 0/100

Prepared using thiazole-4-carboxylic acid

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 2.29-2.40 (m, 2H) 3.05-3.12 (m,2H) 3.49-3.56 (m, 2H) 3.80-4.45 (m, 9H) 5.80-5.86 (m, 1H) 7.60-7.62 (m,1H) 8.04-8.18 (m, 1H) 8.21-8.25 (m, 1H) 8.64-8.68 (m, 1H) 8.79-8.84 (m,1H)

Example 56:{(S)-3-[6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

To a6-(5-chloro-6-methoxy-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(97 g, 0.196 mmol) in CH₂Cl₂ (5 mL) was added the acid chloridetetrahydro-2H-pyran-4-carbonyl chloride (36.7 g, 0.235 mmol) andtriethylamine (0.035 mL, 0.254 mmol) at temperature between 0-10° C. Thereaction mixture was stirred at ˜3° C. for 30 min. The reaction mixturewas concentrated under vacuum. Purification by preparative reverse phaseGilson HPLC and subsequent neutralization of the combined fractions overPL-HCO₃ MP gave{(S)-3-[6-(5-chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone(38 mg, 41% yield) as a white lyophilized powder. ¹H NMR (400 MHz,CDCl₃-d, 298 K) δ ppm 1.56-1.68 (m, 2H) 1.86-2.04 (m, 2H) 2.20-2.40 (m,2H) 2.50-2.72 (m, 1H) 3.05-3.13 (m, 2H) 3.38-4.16 (m, 16H) 5.70-5.78 (m,1H) 7.42-7.45 (m, 1H) 7.78-7.81 (m, 1H) 8.61-8.66 (m, 1H). LCMS:[M+H]⁺=474.2, Rt⁽²⁾=1.52 min.

6-(5-Chloro-6-methoxy-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

(S)-3-[6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (766.2 g, 1.66 mmol) was dissolved in a TFA/CH₂Cl₂(1/2) solution and stirred at rt for 1 h. The reaction mixture wasconcentrated under vacuum, the residue was diluted with CH₂Cl₂, theorganic layer washed with NaOH 1N then brine, dried over Na₂SO₄,filtered and evaporated to give6-(5-chloro-6-methoxy-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine.LCMS: [M+H]⁺=362.1, Rt (3)=1.28 min.

(S)-3-[6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester

X-Phos (0.073 g, 0.154 mmol), Pd₂(dba)₃ (0.100 g, 0.110 mmol), NaOtBu(0.395 g, 4.11 mmol) and 5-bromo-3-chloro-2-methoxy-pyridine (0.732 g,3.29 mmol) were combined and flushed under a stream of argon for 10 min.To this mixture, a solution of(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (Intermediate 7) (2.15 g, 6.72 mmol) in THF (6 mL)was added at rt and the reaction mixture was stirred at 90° C. for 3 h.The reaction was cooled down to rt, EtOAc was added, the mixturefiltered through a celite pad and concentrated under vacuum.Purification by flash chromatography on silica gel (CH₂Cl₂/MeOH 99/1 to95/5) gave(S)-3-[6-(5-chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester as yellow foam (0.766 g, 60% yield). LCMS:[M+H]⁺=462.1, Rt⁽³⁾=1.84 min

Example 57:{(S)-3-[6-(6-Amino-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

To a solution of (S)-tert-butyl3-(6-(6-(bis(tert-butoxycarbonyl)amino)-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1 (120 g, 0.18 mmol) in CH₂Cl₂ (2.0 mL), was added TFA (2.0mL) and the mixture stood at rt for 1 h. Concentrated in vacuo andeluted through an Isolute® SCX-2 cartridge, eluting with methanol, thenwith 2M ammonia in methanol. Basic fractions were concentrated in vacuoto give5-[4-((S)-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]-3-(trifluoromethyl)pyridin-2-yl)amine(61 mg, 90% yield).5-[4-((S)-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]-3-(trifluoromethyl)pyridin-2-yl)amine(30 mg, 0.079 mmol) was dissolved in CH₂Cl₂ (2.0 mL) and was addedsimultaneously portionwise with sat.NaHCO₃(aq) (2.0 mL) to a vigorouslystirring solution of tetrahydro-2H-pyran-4-carbonyl chloride (15 g, 0.10mmol) in CH₂Cl₂ (2.0 mL) at rt. The resulting biphasic mixture wasstirred at rt for 1 h. Diluted with CH₂Cl₂ (10 mL) and the organic layerwas separated by filtering through a phase separation tube andconcentrated in vacuo. Purification by reverse phase Gilson HPLC (MethodA) and subsequent neutralization of the combined fractions by elutionthrough an Isolute® SCX-2 cartridge, eluting with methanol, then with 2Mammonia in methanol. Basic fractions were concentrated in vacuo to give{(S)-3-[6-(6-amino-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanoneas a pale yellow powder (19 mg, 50% yield)¹H NMR (400 MHz, CDCl₃, 298K)δ ppm 1.56-1.72 (m, 2H) 1.87-2.03 (m, 2H) 2.23-2.74 (m, 3H) 3.04-3.14(m, 2H) 3.48-4.13 (m, 12H) 5.15-5.43 (m, 2H, Ar—NH2) 5.73-5.79 (m, 1H)7.55-7.64 (m, 1H) 7.93-8.02 (m, 1H) 8.61-8.67 (m, 1H) LCMS:[M+H]+=397.1, Rt⁽³⁾=1.32 min.

(S)-tert-butyl3-(6-(6-(bis(tert-butoxycarbonyl)amino)-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carboxylate

To a glass vial was added(S)-3-(5,6,7,8-Tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (Intermediate 7) (100 g, 0.312 mmol),imidodicarbonic acid, 2-[5-bromo-3-(trifluoromethyl)-2-pyridinyl]-,1,3-bis(1,1-dimethylethyl) ester (138 g, 0.312 mmol), cesium carbonate(203 g, 0.62 mmol), tris(dibenzylideneacetone)dipalladium(0) (29 g, 0.03mmol), X-Phos (51 g, 0.11 mmol) and anhydrous dioxane (2 mL). The vialwas flushed with a stream of argon for 15 sec and capped. The mixturewas heated with stirring for 1 h at 110° C. and then stirred at roomtemperature for 18 h. Allowed to cool and partitioned between CH₂Cl₂ (10mL) and water (2 mL) and filtered the biphasic mixture through a celitepad. The organic layer was separated by filtering through phaseseparation tube and concentrated in vacuo. Purification by reverse phaseGilson HPLC (Method A) to give (S)-tert-butyl3-(6-(6-(bis(tert-butoxycarbonyl)amino)-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidine-1-carboxylatetrifluoroacetate (as a yellow gum (120 mg, 48% yield). LCMS:[M+H]+=681.5, Rt⁽⁴⁾=1.49 min.

Imidodicarbonic acid, 2-[5-bromo-3-(trifluoromethyl)-2-pyridinyl]-,1,3-bis(1, 1-dimethylethyl) ester

To 5-bromo-3-(trifluoromethyl)pyridin-2-amine (1.72 g, 7.14 mmol),triethylamine (0.995 mL, 7.14 mmol) and 4-dimethylaminoyridine (20 g,0.164 mmol) in CH₂Cl₂ (50 mL) was added di-tert-butyl-dicarbonate (3.89g, 17.84 mmol) and the resulting mixture stirred at room temperature for18 h. Evaporated to dryness in vacuo and triturated in heptane (25 mL)for 72 h. The resulting precipitate was filtered and washed with heptane(10 mL) to give Imidodicarbonic acid,2-[5-bromo-3-(trifluoromethyl)-2-pyridinyl]-, 1,3-bis(1,1-dimethylethyl)ester as a beige solid (2.23 g, 71% yield). ¹H NMR (400 Mhz, CDCl₃,298K) 1.35 (s, 18H) 8.15 (d, 1H) 8.76 (d, 1H) LCMS: [M+H]+=441/443.1,Rt⁽⁴⁾=1.46 min.

Example 58:{3-[6-(6-Methoxy-5-trifluoromethyl-pyrido[4,3-d]pyrimidin-4-yloxy]-azetidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-azetidine-1-carboxylicacid tert-butyl ester (186 g, 0.312 mmol) in CH₂Cl₂ (2.0 mL) was addedTFA (1.0 mL) and the mixture stirred at room temperature for 1 h.Evaporated in vacuo to give4-(azetidin-3-yloxy)-(6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineditrifluoroacetate as a brown gum (110 mg). To a vigorously stirringsolution of tetrahydro-2H-pyran-4-carbonyl chloride (19 g, 0.128 mmol)in CH₂Cl₂ was added simultaneously portionwise sat. NaHCO₃(aq) (2.0 mL)and a solution of4-(azetidin-3-yloxy)-(6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineditrifluoroacetate (60 g, 0.099 mmol) in CH₂Cl₂ (2.0 mL) at rt. Theresulting biphasic mixture was stirred vigorously at rt for 1 h. Dilutedwith CH₂Cl₂ (10 mL), the organic layer separated, dried (MgSO₄),concentrated in vacuo and purified by reverse phase Gilson HPLC (MethodA) and subsequent neutralization of the combined fractions by elutingthrough an Isolute® SCX-2 cartridge, eluting with methanol, then with 2Mammonia in methanol. Basic fractions were concentrated in vacuo to give{3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-azetidin-1-yl}-(tetrahydro-pyran-4-yl)-methanoneas a yellow solid (3.0 mg, 5% yield 2^(nd) step)¹H NMR (400 MHz,DMSO-d6, 298K) δ ppm 1.42-1.67 (m, 4H) 2.90-2.98 (m, 2H) 3.55-3.62 (m,2H) 3.78-4.32 (m, 13H) 4.61-4.69 (m, 1H) 5.42-5.49 (m, 1H) 7.86-7.90 (m,1H) 8.22-8.26 (m, 1H) 8.58-8.62 (s, 1H) LCMS: [M+H]+=494.6, Rt⁽⁷⁾=0.98min.

3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-azetidine-1-carboxylicacid tert-butyl ester

To a glass vial was added3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-azetidine-1-carboxylicacid tert-butyl ester (110 g, 0.359 mmol),5-Bromo-2-methoxy-3-(trifluoromethyl)pyridine (92 g, 0.359 mmol), cesiumcarbonate (234 g, 0.718 mmol), tris(dibenzylideneacetone)dipalladium(0)(33 g, 0.036 mmol), X-Phos (58 g, 0.122 mmol) and anhydrous dioxane (2.0mL). The vial was flushed with a stream of argon for 15 sec and capped.The mixture was heated with stirring for 1.5 h at 110° C. and thenstirred at room temperature for 18 h. Diluted with CH₂Cl₂ (50 mL),filtered through a celite pad and concentrated in vacuo. Purified byreverse phase Gilson HPLC (Method A) to give the3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-azetidine-1-carboxylicacid tert-butyl ester trifluoroacetate as a brown gum (186 mg, 87%yield) LCMS: [M+H]+=482.3, Rt⁽⁷⁾=1.56 min.

3-(5,6,7,8-Tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-azetidine-1-carboxylicacid tert-butyl ester

To a solution of3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-azetidine-1-carboxylicacid tert-butyl ester (425 g, 1.07 mmol) in MeOH (20 mL) was added 20%palladium hydroxide on carbon (90 mg) then ammonium formate (473 g, 7.51mmol) and the mixture heated at reflux for 1 h. The reaction mixture wasallowed to cool and filtered through a celite pad, washing with MeOH (20mL) then CH₂Cl₂ (20 mL). The filtrate was evaporated in vacuo andpurified by flash chromatography on silica gel with CH₂Cl₂/MeOH/0.88NH₄OH, 100/0/0 to 90/10/1 to give3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-azetidine-1-carboxylicacid tert-butyl ester as a yellow gum (220 mg, 67% yield) LCMS:[M+H]+=307.3, Rt⁽⁴⁾=0.81 min.

3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-azetidine-1-carboxylicacid tert-butyl ester 3-Hydroxy-azetidine-1-carboxylic acid tert-butylester (217 g, 1.25 mmol) was dissolved under argon in THF (10 mL) andNaH (58 g, 1.44 mmol) was added. The resulting suspension was stirred atrt under argon for 15 min following by the addition of a solution of6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3d]pyrimidine (250 g,0.963 mmol). The reaction mixture was stirred at rt for 18 h, quenchedwith water (20 mL) and diluted with CH₂Cl₂. The organic layer wasfiltered through a phase separation tube and concentrated in vacuo.Purification by flash chromatography on silica gel with heptane/CH₂Cl₂,50/50 to 0/100 then EtOAc to give3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-azetidine-1-carboxylicacid tert-butyl ester as a yellow gum (425 mg, 111% yield) LCMS:[M+H]+=397.4, Rt⁽⁴⁾=0.98 min.

Example 59 was prepared according the general procedure described inscheme 2.

Example 59:{(S)-3-[6-(2-Methoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Step 4

A mixture of 5-bromo-2-methoxy-pyrimidine (0.218 mmol), X-Phos (28.4 g,0.060 mmol), Pd₂(dba)₃ (18.2 g, 0.020 mmol) and Cs₂CO₃ (129 g, 0.397mmol) was flushed with argon before the addition of a solution of(tetrahydro-pyran-4-yl)-[(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-methanonein dioxane (2 mL). The reaction mixture was heated at 120° C. for 1 h ina sealed vial, cooled down to rt and filtered over Hyflo, The recoveredorganic phase was washed with NaHCO₃ and brine, dried over Na₂SO₄,filtered and concentrated. Purification by preparative reverse phaseGilson HPLC and neutralization of the combined fractions by passingthrough a SCX-2 cartridge (The cartridge was washed with acetonitrile,CH₂Cl₂ and MeOH, then a solution of NH₃ in MeOH 3.5 N was used toreleased the expected product) gave{(S)-3-[6-(2-methoxy-pyrimidin-5-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone(18.7 mg, 21% yield)¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 1.62-1.70 (m,2H) 1.87-2.01 (m, 2H) 2.20-2.41 (m, 2H) 2.49-2.71 (m, 1H) 3.07-3.19 (m,2H) 3.37-4.19 (m, 16H) 5.76 (m, 1H) 8.32 (s, 2H) 8.65-8.67 (m, 1H).LCMS: [M+H]+=441.2, Rt⁽¹⁾=1.12 min.

(Tetrahydro-pyran-4-yl)-[(S)-3-(5, 6, 7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-methanone

Step 3

A solution of[(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-(tetrahydro-pyran-4-yl)-methanone(10.9 g, 25.8 mmol) was dissolved in methanol (300 mL) and Pd(OH)₂ onCarbon (2 g, 14.24 mmol) and ammonium formate (3.35 g, 51.6 mmol) wereadded. The reaction mixture was refluxed for 2 h. The reaction wascooled down to rt, the reaction mixture was filtered and evaporatedunder high vacuum for 2 h to yield(tetrahydro-pyran-4-yl)-[(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-methanone(8.45 g, 95% yield) as a light yellow foam. ¹H NMR (400 MHz, DMSO-d6,298K) δ ppm 1.44-1.67 (m, 4H) 2.08-2.32 (m, 2H) 2.55-2.83 (m, 3H) 2.96(t, 2H) 3.22-3.96 (m, 11H) 5.53-5.68 (m, 1H) 8.49-8.59 (m, 1H). LCMS:[M+H]+=333.5, Rt⁽⁶⁾=1.24 min.

[(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-(tetrahydro-pyran-4-yl)-methanone

Step 2

To a solution of6-benzyl-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(420 g, 1.35 mmol) in 4 mL of CH₂Cl₂ was addedtetrahydro-pyran-4-carbonyl chloride (0.210 mL, 1.637 mmol) and Et₃N(0.380 mL, 2.73 mmol). The reaction mixture was stirred at roomtemperature for 30 min then was quenched with H₂O, extracted withCH₂Cl₂, filtered and evaporated under vacuum. Purification byflash-chromatography on silica gel (CH₂Cl₂/MeOH 95/5) gave[(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-(tetrahydro-pyran-4-yl)-methanone(420 mg, 73% yield) as a yellow foam. ¹H NMR (400 MHz, DMSO-d6, 298K) δppm 1.37-1.64 (m, 4H) 1.95-2.29 (m, 2H) 2.56-2.83 (m, 4H) 3.28-3.91 (m,13H) 5.54-5.68 (m, 1H) 7.24-7.36 (m, 5H) 8.54-8.59 (m, 1H). LCMS:[M+H]+=423.6, Rt⁽⁷⁾=0.68.

6-Benzyl-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

Step 1

To a solution of(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (560 g, 1.364 mmol) in 2 mL of CH₂Cl₂ was addedTFA (1.576 mL, 20.46 mmol). The reaction mixture was stirred at rt for 1h, concentrated and then eluted through an Isolute SCX-2 cartridge (10g) to remove excess TFA with (i) MeOH (ii) NH₃/MeOH and the basicfraction evaporated in vacuum to give6-benzyl-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(420 mg, quantitative yield) as a yellow gum. LCMS: [M+H]+=311.2,Rt⁽³⁾⁼0.11.

Examples 60-62 were prepared using procedures analogous to those used inExample 59 using appropriate starting materials.

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 60

1.27 460.2

Name:[(S)-3-(6-Quinolin-3-yl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy-pyrrolidin-1-yl]-(tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 3-bromo-quinoline

¹H NMR (400 MHz, CDCl₃-d, 298K) δ ppm 1.60-1.72 (m, 3H) 1.89-2.04 (m,2H) 2.24-2.46 (m, 2H) 2.53-2.73 (m, 1H) 3.08-3.21 (m, 2H) 3.39-3.52 (m,2H) 3.67-4.11 (m, 8H) 4.20-4.40 (m, 2H) 5.73-5.83 (m, 1H) 7.47-7.62 (m,3H) 7.73-7.80 (m, 1H) 8.02-8.14 (m, 1H) 8.62-8.68 (m, 1H) 8.86-8.91 (m,1H).

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 61

1.17 508.7

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using intermediate 1

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.50-1.66 (m, 4H) 2.07-2.46 (m,2H) 2.60-2.80 (m, 1H) 2.88-2.97 (m, 2H) 3.30-3.95 (m, 13H) 4.08-4.23 (m,2H) 5.59-5.74 (m, 1H) 7.79-7.85 (m, 1H) 8.16-8.23 (m, 1H) 8.60-8.65 (m,1H)

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 62

2.11 494.2

Name:1-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-3,3-dimethyl-butan-1-one

Purification method: Reverse phase method A

Prepared using step 2 of example 59 and 3,3-dimethyl-butyryl chloridefollowed by steps 3-4 of example 59 using intermediate 1

¹H NMR (400 MHz, CDCl₃, 298K) δ ppm 1.00-1.12 (m, 9H) 2.13-2.35 (m, 4H)3.08-3.15 (m, 2H) 3.45-3.93 (m, 6H) 4.02 (s, 3H) 4.03-4.15 (m, 2H)5.72-5.79 (m, 1H) 7.57-7.62 (m, 1H) 8.03-8.07 (m, 1H) 8.64-8.69 (m, 1H)

Example 63 was prepared according the general procedure described inscheme 2.

Example 63:1-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-propan-1-one

Step 3

1-[(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-propan-1-one(47.8 g, 0.173 mmol), X-Phos (28 g, 0.059 mmol) and Pd₂(dba)₃.CHCl₃(17.90 mg, 0.017 mmol) were combined and flushed with argon duringseveral min before addition of degassed dioxane.5-Bromo-2-methoxy-3-trifluoromethyl-pyridine (intermediate 1) (54.5 g,0.213 mmol) and Cs₂CO₃ (113 g, 0.346 mmol) were then added to thereaction mixture and the resulting mixture flushed with argon and heatedat 150° C. for 30 min. in a sealed tube. The reaction mixture was cooledto rt, filtered over Hyflo and evaporated. Purification by preparativereverse phase Gilson HPLC and subsequent neutralization of the combinedfractions over PL-HCO₃ MP gave1-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-propan-1-one(26 mg, 33% yield)¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 0.94-1.00 (m,3H) 2.05-2.17 (m, 4H) 2.95-3.0 (m, 2H) 3.45-3.97 (m, 9H) 4.07-4.11 (m,2H) 5.58-5.72 (m, 1H) 7.81-7.86 (m, 1H) 8.18-8.23 (m, 1H) 8.62 (s, 1H).MS: [M+H]+=452.2, Rt⁽¹⁾=1.74 min.

1-[(S)-3-(5,6,7,8-Tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-propan-1-one

Step 2

Pd(OH)₂ (150 g, 1.070 mmol) was put into a round flask and flushed underargon for 5 minutes. A solution of1-[(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-propan-1-one(560 g, 1.528 mmol) in 22 mL of MeOH was added followed by ammoniumformate (482 g, 7.64 mmol). The reaction mixture was stirred underreflux (70° C.) for 2 h. The mixture was filtered over a pad of celiteand dried under high vacuum to give1-[(S)-3-(5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-propan-1-one.No further purification (m=420 mg, quantitative yield). MS: [M+H]+=277.5Rt⁽⁶⁾=0.71 min.

1-[(S)-3-(6-Benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-propan-1-one

Step 1

To a solution of 1-((S)-3-hydroxy-pyrrolidin-1-yl)-propan-1-one(intermediate 2) (358 g, 2.503 mmol) in 5 mL of THF was added NaH (108g, 2.70 mmol) under Ar. The mixture was stirred at rt for 15 min, then6-benzyl-4-chloro-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine (500 g,1.925 mmol) and 5 mL of THF were added and stirred at rt for 5 h. Thereaction was quenched with H₂O and extracted with ethylacetate, the org.layer was washed with brine, dried over MgSO₄, filtered and evaporatedto dryness. The residue was purified by flash-chromatography using IscoCompanion system (12 g of SiO₂) CH₂Cl₂/MeOH (95/5). The collectedfractions were combined, evaporated and dried over high vacuum to give1-[(S)-3-(6-benzyl-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)-pyrrolidin-1-yl]-propan-1-one.(m=560 mg, yield 78%) MS: [M+H]+=367.6, Rt⁽⁷⁾=0.64 min.

Example 64 was prepared using procedures analogous to those used inExample 63 using appropriate starting materials.

Example Rt⁽¹⁾ (min.) MS: [M + H]+ 64

1.46 409.2

Name:2-Methoxy-5-[4-((S)-1-propionyl-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]-nicotinonitrile

Purification method: Reverse phase method A

Prepared using step 3 of example 59 and 5-bromo-2-methoxynicotinonitrile

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 0.95-1.02 (m, 3H) 2.10-2.35 (m,5H) 2.89-2.98 (m, 2H) 3.40-3.90 (m, 5H) 3.93 (s, 3H) 4.16 (s, 2H)5.58-5.71 (m, 1H) 8.08-8.10 (m, 1H) 8.24-8.28 (m, 1H) 8.61 (m, 1H).

Examples 65:6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-1-pyridin-2-yl-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

To a glass vial was added6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidinedihydrochloride (prepared using step 1 of example 91 from intermediate13) (75 g, 0.16 mmol), 2-bromopyridine (1 mL, 10.25 mmol) andN,N-diisopropylethylamine (0.14 mL, 0.80 mmol). The vial was capped andthe mixture heated in the microwave at 160° C. for 20 min. Purificationby reverse phase Gilson HPLC (Method A) and subsequent neutralization ofthe combined fractions over PL-HCO₃ MP to give6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-1-pyridin-2-yl-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineas a light brown solid (19 mg, 25% yield)¹H NMR (400 MHz, DMSO-d6, 298K)δ ppm 2.24-2.44 (m, 2H) 2.92 (t, 2H) 3.47-3.69 (m, 5H) 3.77-3.85 (m, 1H)3.88-3.93 (m, 3H) 4.12-17 (m, 2H) 5.73-5.81 (m, 1H) 6.40-6.52 (d, 1H)6.56-6.58 (m, 1H) 7.43-7.54 (m, 1H) 7.77-7.84 (m, 1H) 8.02-8.09 (m, 1H)8.13-8.20 (m, 1H) 8.61-8.66 (m, 1H) LCMS: [M+H]+=473.0, Rt⁽⁴⁾=0.85 min.

Examples 66:6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-1-pyrimidin-2-yl-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine

To a glass vial was added6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidinedihydrochloride (prepared using step 1 of example 91 from intermediate13) (75 g, 0.16 mmol), 2-bromopyrimidine (55 mg, 0.342 mmol) andN,N-diisopropylethylamine (0.15 mL, 0.85 mmol). The vial was capped andthe mixture heated in the microwave at 160° C. for 20 min. Purificationby reverse phase Gilson HPLC (Method A) and subsequent neutralization ofthe combined fractions over PL-HCO₃ MP to give6-(6-methoxy-5-trifluoromethyl-pyridin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidineas a brown solid (17 mg, 21% yield)¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm2.23-2.43 (m, 2H) 2.85-2.99 (t, 2H) 3.22-3.94 (m, 9H) 4.08-4.27 (m, 2H)5.70-5.80 (m, 1H) 6.56-6.66 (t, 1H) 7.76-7.87 (m, 1H) 8.12-8.27 (m, 1H)8.28-8.42 (m, 2H) 8.59-8.68 (m, 1H) LCMS: [M+H]+=474.2, Rt⁽¹⁾=1.91 min.

Example 67 was prepared according the general procedure described inscheme 4

Example 67:1-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-propan-1-one

To a solution of(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 24) (13.4 g, 27.1 mmol) in CH₂Cl₂(100 mL), was added TFA (41.8 mL) and the mixture stirred at rt for 1 h.Concentrated in vacuo and partitioned between 2M NaOH(aq) (300 mL) andCH₂Cl₂ (200 mL). The organic phase was separated and the aqueous phaseextracted with CH₂Cl₂ (2×200 mL). The organic phases were combined,dried (MgSO₄) and evaporated in vacuo to give a brown foam. The foam wasdissolved in CH₂Cl₂ (50 mL) and was added simultaneously portionwisewith sat.NaHCO₃(aq) (50 mL) to a vigorously stirring solution ofpropionyl chloride (2.63 g, 28.5 mmol) in CH₂Cl₂ (50 mL) at rt. Theresulting biphasic mixture was stirred at rt for 1 h. Further propionylchloride (0.566 g, 6.12 mmol) was added and continued stirringvigorously for 20 min. The organic layer was separated and the aqueouslayer extracted with CH₂Cl₂ (100 mL). The organic layers were combined,dried (MgSO₄) and concentrated in vacuo to give a brown gum. The gum wasstirred in EtOAc (100 mL) and the resulting solid filtered (9.4 g). Themother liquors were concentrated in vacuo and purified by columnchromatography through a Biotage® amino silica gel eluting withEtOAc/MeOH, 100/0 to 90/10 to give a yellow foam which was then stirredin EtOAc (20 mL) and the resulting solid filtered (870 mg). Both batchesof solids were combined and stirred in refluxing EtOAc (50 mL) for 1 h.Filtered to give1-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-propan-1-oneas a colourless solid (9.42 g, 76% yield). ¹H NMR (400 MHz, DMSO-d6,298K) δ ppm 0.95-1.05 (m, 3H) 1.87-2.32 (m, 4H) 2.77-2.86 (m, 2H)3.25-3.88 (m, 6H) 3.93 (s, 3H) 3.98 (s, 2H) 4.55-4.80 (m, 1H) 6.70-6.80(m, 1H, N—H) 7.86-7.92 (m, 1H) 8.27-8.33 (m, 1H) 8.33-8.37 (m, 1H) LCMS:[M+H]+=451.0, Rt⁽⁶⁾=1.49 min.

Alternative Synthesis for Example 67

A solution of(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 24) (29.04 g, 58.73 mmol) in2-Me-THF (100 mL) was dropwise added into aqueous HCl solution (150 mL,31%) over 15 min. The reaction mixture was partitioned between water(300 mL) and isopropyl acetate (100 mL) and the upper organic phase wasdiscarded. The aqueous phase was partitioned between 25% NaOH (aq) (200g) and 2-Me-THF (200 mL), and the organic phase was collected and dried.Triethylamine (16.32 mL, 117.48 mmol) was added into the organic phasefollowed by dropwise addition of propionyl chloride (6.0 g, 64.6 mmol)at 0° C. The resulting mixture was stirred at 0° C. for 1 h. Thereaction mixture was washed with water (110 mL) and the resultingorganic phase was concentrated in vacuo to give a brown gum. The residuewas recrystallized with isopropanol and methyl tert-butyl ether to give1-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-propan-1-oneas a colourless solid (17.2 g, 65% yield).

Crystallization of Example 67 by Heating in Acetonitrile/Water

2.0 g of Example 67 (4.440 mol) were dissolved in 10 mL of acetonitrileand 0.5 mL of water at 75° C. The solution was allowed to cool down tort within 30 min resulting in a suspension. The mixture was stirred for16 h at rt. The crystals were collected by filtration. The filter cakewas washed 2 times with 1 mL of acetonitrile and afterwards dried for 16h at 24° C. and ca. mbar vacuum. Elementary analysis of the materialshowed a waterless form.

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 67 anhydrous form Method X1):

2-Theta in deg Intensity in % 7.9 31 9.6 88 11.5 29 13.4 8 15.2 7 15.9100 16.8 57 17.6 9 18.7 20 20.0 8 20.6 40 22.0 32 22.4 53 22.7 26 23.417 23.9 23 24.5 41 25.1 20 25.8 13 26.7 31

Preparation of Phosphate Salt of Example 67

2.0 g of Example 67 (4.440 mol) were dissolved in 10 mL of acetonitrileand 0.5 mL of water at 75° C. 512 mg of ortho-phosphoric acid 85% (4.440mol) were added at 70° C. Crystallization occurs quickly at 70° C. Thesuspension was allowed to cool down to rt within 30 min. The suspensionwas diluted with 10 ml acetonitrile and stirred for 16 h at rt. Thecrystals were collected by filtration. The filter cake was washed 3times with 1 mL of acetonitrile and afterwards dried for 16 h at 24° C.and ca. 10 mbar vacuum. Elementary analysis of the phosphate salt showeda 1:1 (waterless) form

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 67 phosphate salt (Method X1):

2-Theta in deg Intensity in % 5.2 51 9.8 56 10.3 19 11.6 100 14.9 1415.5 48 15.9 11 16.6 65 19.5 54 20.7 62 21.5 10 22.1 21 23.3 57 25.8 1826.4 29 27.2 20 28.2 13

Preparation of hydrochloride salt of Example 67

2.0 g of Example 67 (4.440 mol) were dissolved in 20 mL of acetonitrileand 1.0 mL of water at 70° C. 459 mg of hydrochloric acid 37% (4.440mol) were added at 70° C. Crystallization occurs quickly at 70° C. Thesuspension was allowed to cool down to rt within 30 min and stirred for16 h at rt. The crystals were collected by filtration. The filter cakewas washed 3 times with 1 mL of acetonitrile and afterwards dried for 16h at 24° C. and ca. 10 mbar vacuum. Elementary analysis of the HCl saltshowed a 1:1 (waterless) form

List of most significant peaks from X-ray Powder Diffraction Pattern ofExample 67 hydrochloride salt (Method X1):

2-Theta in deg Intensity in % 5.6 100 11.0 18 11.3 42 11.8 12 14.7 3317.1 13 18.7 19 19.4 29 22.0 23 22.6 28 23.1 50 23.7 28 24.9 29 25.5 15

Preparation of Hippurate Salt of Example 67

0.4 g of Example 67 (0.888 mmol) were dissolved in 8 mL of acetonitrileand 0.2 mL of water at 70° C. 167 mg of hippuric acid (0.888 mmol) wereadded at 70° C. The solution was allowed to cool down to rt within 30min. Crystallization occurs at 40° C. The suspension was stirred for 16h at rt. The crystals were collected by filtration. The filter cake waswashed 3 times with 1 mL of acetonitrile and afterwards dried for 16 hat 50° C. and ca. 10 mbar vacuum List of most significant peaks fromX-ray Powder Diffraction Pattern of Example 67 hippurate salt (MethodX1):

2-Theta in deg Intensity in % 5.2 76 7.5 100 10.3 60 10.9 63 11.8 9 13.116 16.1 44 16.7 26 17.7 49 18.4 38 21.2 49 23.2 74 24.2 67 26.2 28

Examples 68-69 were prepared using procedures analogous to those used inexample 67 using appropriate starting materials.

Rt⁽¹⁾ MS: Example (min.) [M + H]+ 68

1.26 507.2

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase HPLC Method A

Prepared using tetrahydro-pyran-4-carbonyl chloride

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.48-1.67 (m, 4H) 1.88-2.35 (m,2H) 2.59-2.87 (m, 3H) 3.26-4.03 (m, 15H) 4.56-4.83 (m, 1H) 6.82-6.92 (m,1H, N—H) 7.86-7.90 (m, 1H) 8.26-8.32 (m, 1H) 8.37-8.42 (m, 1H)

Rt⁽¹⁾ MS: Example (min.) [M + H]+ 69

1.06 464.2

Name:2-Methoxy-5-{4-[(S)-1-(tetrahydro-pyran-4-carbonyl)-pyrrolidin-3-ylamino]-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl}-nicotinonitrile

Purification method: Reverse phase HPLC Method A

Prepared using intermediate 25 and tetrahydro-pyran-4-carbonyl chloride

¹H NMR (400 MHz, MeOH-d4, 298K) δ ppm 1.59-1.86 (m, 4H) 2.07-2.47 (m,2H) 2.75-2.98 (m, 3H) 3.44-4.13 (m, 15H) 4.64-5.24 (m, 1H, signal maskedby water peak) 7.94-7.99 (m, 1H) 8.20-8.26 (m, 1H) 8.33-8.39 (m, 1H)

Example 70:1-(4-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanone

(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester (intermediate 24) (160 g, 0.32 mmol) was dissolvedin CH₂Cl₂ (2.0 mL) and TFA (1.0 mL) added. The resulting mixture wasstirred at room temperature for 1 h then evaporated in vacuo to give[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yl]-(S)-pyrrolidin-3-yl-amineditrifluoroacetate as a brown gum (160 mg), which was used withoutfurther purification. To[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yl]-(S)-pyrrolidin-3-yl-amineditrifluoroacetate (40 g, 0.06 mmol) was added1-acetylpiperidine-4-carboxylic acid (12 g, 0.07 mmol)),N,N-diisopropylethylamine (0.05 mL, 0.26 mmol), CH₂Cl₂ (3.0 mL) and thenHBTU (29 g, 0.08 mmol). The mixture was allowed to stir at roomtemperature for 18 h and then partitioned between CH₂Cl₂ (10 mL) andwater (5 mL). The organic phase was filtered through a phase separationtube and evaporated in vacuo. Purification by reverse phase Gilson HPLC(Method A) and subsequent neutralization of the combined fractions byelution through an Isolute® SCX-2 cartridge, eluting with methanol, thenwith 2M ammonia in methanol. Basic fractions were concentrated in vacuoto give1-(4-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanoneas a pale yellow solid (19 mg, 50% yield for 2^(nd) step)¹H NMR (400MHz, DMSO-d6, 298K) δ ppm 1.20-1.70 (m, 4H) 1.79-2.35 (m, 5H) 2.53-2.85(m, 4H) 3.04-3.14 (m, 1H) 3.35-4.79 (m, 14H) 6.80-6.87 (m, 1H, N—H)7.87-7.91 (m, 1H) 8.26-8.31 (m, 1H) 8.35-8.41 (m, 1H) LCMS:[M+H]+=548.2, Rt⁽¹⁾=1.22 min.

Examples 71-80 were prepared using procedures analogous to those used inexample 70 using appropriate starting materials

Rt⁽¹⁾ MS: Example (min.) [M + H]+ 71

1.40 535.3

Name:(2,2-Dimethyl-tetrahydro-pyran-4-yl)-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase HPLC Method A

Prepared using 2,2-dimethyl-tetrahydro-pyran-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.05-1.20 (m, 6H) 1.30-1.58 (m,4H) 1.86-2.35 (m, 2H) 2.70-2.90 (m, 3H) 3.34-4.03 (m, 13H) 4.55-4.80 (m,1H) 6.67-6.76 (m, 1H, N—H) 7.86-7.89 (m, 1H) 8.26-8.31 (m, 1H) 8.32-8.37(m, 1H)

Example 72

Rt⁽¹⁾ (min.) 1.22 MS: [M + H]+ 490.1

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-oxazol-5-yl-methanone

Purification method: Reverse phase HPLC Method A

Prepared using oxazole-5-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.89-2.39 (m, 2H) 2.78-2.86 (m,2H) 3.50-4.20 (m, 11H) 4.65-4.84 (m, 1H) 6.75-6.83 (m, 1H, N—H)7.75-7.83 (m, 1H) 7.86-7.92 (m, 1H) 8.26-8.32 (m, 1H) 8.35-8.38 (m, 1H)8.55-8.60 (m, 1H)

Example 73

Rt⁽¹⁾ (min.) 1.38 (Isomer 1) MS: [M + H]+ 535.2 (Isomer 1) Example 74

1.48 (Isomer 2) Cis/trans not assigned 535.2 (Isomer 2)

Name:((S)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)((1R,4R)-4-methoxycyclohexyl)methanone

((S)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)((1R,4S)-4-methoxycyclohexyl)methanone

Purification method: Reverse phase HPLC Method A

Prepared using 4-methoxy-cyclohexanecarboxylic acid (mixture ofcis/trans)

¹H NMR (Isomer 1 cis/trans not assigned) (400 MHz, DMSO-d6, 298K) δ ppm1.04-1.47 (m, 4H) 1.64-2.45 (m, 7H) 2.77-2.86 (m, 2H) 3.00-3.77 (m, 10H)3.87-4.03 (m, 5H) 4.53-4.80 (m, 1H) 6.67-6.78 (m, 1H, N—H) 7.85-7.91 (m,1H) 8.26-8.32 (m, 1H) 8.33-8.38 (m, 1H)

¹H NMR (Isomer 2 cis/trans not assigned) (400 MHz, DMSO-d6, 298K) δ ppm1.32-1.48 (m, 4H) 1.55-2.50 (m, 7H) 2.78-2.84 (m, 2H) 3.01-3.77 (m, 10H)3.87-4.03 (m, 5H) 4.53-4.80 (m, 1H) 6.67-6.78 (m, 1H, N—H) 7.85-7.91 (m,1H) 8.26-8.32 (m, 1H) 8.33-8.38 (m, 1H)

Example 75

Rt⁽¹⁾ (min.) 1.18 (Isomer 1) MS: [M + H]+ 521.2 (Isomer 1) Example 76

1.29 (Isomer 2) Cis/trans not assigned 521.2 (Isomer 2)

Name: ((1S,4R)-4-hydroxycyclohexyl)((S)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone

((1R,4S)-4-hydroxycyclohexyl)((S)-3-(6-(6-methoxy-5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl)methanone

Purification method: Reverse phase HPLC Method A

Prepared using 4-hydroxy-cyclohexanecarboxylic acid (mixture ofcis/trans)

¹H NMR (Isomer 1 cis/trans not assigned) (400 MHz, DMSO-d6, 298K) δ ppm1.06-1.44 (m, 4H) 1.57-2.86 (m, 9H) 3.01-3.76 (m, 7H) 3.88-4.03 (m, 5H)4.50-4.78 (m, 2H) 6.68-6.78 (m, 1H, N—H) 7.86-7.91 (m, 1H) 8.26-8.32 (m,1H) 8.33-8.39 (m, 1H)

¹H NMR (Isomer 1 cis/trans not assigned) (400 MHz, DMSO-d6, 298K) δ ppm1.28-1.52 (m, 4H) 1.59-2.85 (m, 9H) 3.03-3.83 (m, 8H) 3.88-4.03 (m, 5H)4.55-4.79 (m, 1H) 6.68-6.77 (m, 1H, N—H) 7.85-7.91 (m, 1H) 8.26-8.32 (m,1H) 8.33-8.39 (m, 1H)

Example 77

Rt⁽¹⁾ (min.) 0.89 MS: [M + H]+ 449.2

Name-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-(1-methyl-1H-imidazol-4-yl)-methanone

Purification method: Reverse phase HPLC Method A

Prepared using intermediate 20 and 1-methyl-1H-imidazole-4-carboxylicacid

¹H NMR (400 MHz, MeOH-d4, 298K) δ ppm 2.07-2.44 (m, 5H) 2.86-2.95 (m,2H) 3.44-4.43 (m, 14H) 4.77-4.87 (m, 1H) 7.44-7.48 (m, 1H) 7.60-7.70 (m,2H) 7.72-7.79 m, 1H) 8.32-8.41 (m, 1H)

Example 78

Rt⁽¹⁾ (min.) 0.99 MS: [M + H]+ 436.2

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-oxazol-5-yl-methanone

Purification method: Reverse phase HPLC Method A

Prepared using intermediate 20 and oxazole-5-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.95-2.35 (m, 5H) 2.74-2.83 (m,2H) 3.35-4.20 (m, 11H) 4.62-4.83 (m, 1H) 6.73-6.81 (m, 1H) 7.44-7.49 (m,1H) 7.75-7.85 (m, 2H) 8.33-8.38 (m, 1H) 8.54-8.59 (m, 1H)

Example 79

Rt⁽¹⁾ (min.) 1.01 MS: [M + H]+ 436.2

Name:{(S)-3-[6-(6-Methoxy-1-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-oxazol-4-yl-methanone

Purification method: Reverse phase HPLC Method A

Prepared using intermediate 20 and oxazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.93-2.35 (m, 5H) 2.75-2.82 (m,2H) 3.38-4.27 (m, 11H) 4.61-4.78 (m, 1H) 6.74-6.80 (m, 1H) 7.45-7.49 (m,1H) 7.78-7.84 (m, 1H) 8.32-8.37 (m, 1H) 8.47-8.53 (m, 1H) 8.61-8.66 (m,1H)

Example 80

Rt⁽¹⁾ (min.) 1.18 MS: [M + H]+ 481.3

Name:(2,2-Dimethyl-tetrahydro-pyran-4-yl)-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-methanone

Purification method: Reverse phase HPLC Method A

Prepared using intermediate 20 and2,2-dimethyl-tetrahydro-pyran-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.05-1.22 (m, 6H) 1.30-1.58 (m,4H) 1.90-2.29 (m, 5H) 2.75-2.85 (m, 3H) 3.35-3.77 (m, 7H) 3.82 (s, 3H)3.87-3.97 (m, 3H) 4.54-4.79 (m, 1H) 6.66-6.75 (m, 1H) 7.47 (d, 1H) 7.81(d, 1H) 8.35 (d, 1H)

Example 81:1-{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-propan-1-one

To a solution of(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidine-1-carboxylicacid tert-butyl ester trifluoroacetate (intermediate 20) (60 g, 0.11mmol) in CH₂Cl₂ (2.0 mL), was added TFA (2.0 mL) and the mixture stirredat rt for 1 h. Concentrated in vacuo to give[6-(6-methoxy-5-methylpyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-yl)]-(S)-pyrrolidin-3-yl)amineditrifluoroacetate (60 mg).[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yl]-(S)-pyrrolidin-3-yl)amineditrifluoroacetate (30 g, 0.053 mmol) was dissolved in CH₂Cl₂ (2.0 mL)and was added simultaneously portionwise with sat.NaHCO₃(aq) (2.0 mL) toa vigorously stirring solution of propionyl chloride (7 g, 0.07 mmol) inCH₂Cl₂ (2.0 mL) at rt. The resulting biphasic mixture was stirred at rtfor 45 min. Diluted with CH₂Cl₂ (10 mL) and sat.NaHCO₃(aq) (2.0 mL). Theorganic layer was separated by filtering through phase separation tubeand concentrated in vacuo. Purification by reverse phase Gilson HPLC(Method A) and subsequent neutralization of the combined fractions byelution through an Isolute® SCX-2 cartridge, eluting with methanol, thenwith 2M ammonia in methanol. Basic fractions were concentrated in vacuogave1-{(S)-3-[6-(6-methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-propan-1-oneas a colourless powder (7 mg, 21% yield)¹H NMR (400 MHz, MeOH-d4, 298K)δ ppm 1.20-1.28 (m, 3H) 2.04-2.44 (m, 7H) 2.88-2.94 (m, 2H) 3.48-4.04(m, 11H) 4.73-4.88 (m, 1H) 7.44-7.48 (m, 1H) 7.73-7.77 (m, 1H) 8.34-8.38(m, 1H) LCMS: [M+H]+=397.1, Rt⁽³⁾=1.32 min.

Examples 82-83 were prepared using procedures analogous to those used inExample 81 using appropriate starting materials.

Example 82

Rt⁽¹⁾ (min.) 1.13 MS: [M + H]+ 473.2

Name:{(S)-3-[6-(5-Chloro-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase HPLC Method A

Prepared using intermediate 21 and tetrahydro-pyran-4-carbonyl chloride¹H NMR (400 MHz, MeOH-d4, 298K) δ ppm 1.58-1.87 (m, 4H) 2.04-2.45 (m,2H) 2.73-2.96 (m, 3H) 3.39-4.14 (m, 15H) 4.71-4.90 (m, 1H) 7.67-7.74 (m,1H) 7.88-7.93 (m, 1H) 8.34-8.39 (m, 1H)

Example 83

Rt⁽¹⁾ (min.) 0.86 MS: [M + H]+ 453.6

Name:{(S)-3-[6-(6-Methoxy-5-methyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Reverse phase HPLC Method A then Method C

Prepared using intermediate 20 and tetrahydro-pyran-4-carbonyl chloride

¹H NMR (400 MHz, MeOD-d6, 298K) δ ppm 1.57-1.88 (m, 4H) 2.04-2.45 (m,5H) 2.73-2.96 (m, 3H) 3.37-4.12 (m, 15H) 4.73-4.88 (m, 1H) 7.45-7.48 (m,1H) 7.73-7.77 (m, 1H) 8.36-8.39 (m, 1H)

Example 84:(Tetrahydro-pyran-4-yl)-{(S)-3-{6-(5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl}-methanone

To6-(5-(trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ol(intermediate 19) (59 mg) in acetonitrile (1.0 ml) was added BOP (114 g,0.258 mmol) and DBU (0.060 ml, 0.398 mmol). The resulting solution wasstood at RT for 1 min then added[(S)-3-Amino-pyrrolidin-1-yl-(tetrahydro-pyran-4-yl)-methanone(intermediate 5) (79 mg, 0.398 mmol) in acetonitrile (1.0 ml) and heatedthe mixture at 85° C. for 25 h. The reaction mixture was evaporated invacuo and purified by reverse phase Gilson HPLC and subsequentneutralization of the combined fractions by eluting through an Isolute®SCX-2 cartridge, eluting with methanol, then with 2M ammonia inmethanol. Basic fractions were concentrated in vacuo gave crude titlecompound which was further purified by flash chromatography on silicagel with EtOAc/MeOH 100/0 to 80/20 to give(Tetrahydro-pyran-4-yl)-{(S)-3-{6-(5-(trifluoromethyl)pyridin-3-yl)-5,6,7,8-tetrahydropyrido[4,3-d]pyrimidin-4-ylamino)pyrrolidin-1-yl}-methanone(19 mg, 6% yield) as a colourless solid. ¹H NMR (400 MHz, DMSO-d6, 298K)δ ppm 1.47-1.69 (m, 4H) 1.83-2.37 (m, 2H) 2.58-2.89 (m, 3H) 3.23-4.20(m, 12H) 4.56-4.82 (m, 1H) 6.75-6.89 (m, 1H, N—H) 7.68-7.79 (m, 1H)8.28-8.42 (m, 2H) 8.74-8.83 (m, 1H). LCMS: [M+H]+=477.6, Rt⁽⁷⁾=0.84 min.

Example 85:{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(4-methyl-piperazin-1-yl)-methanone

To a solution of6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(prepared using step 1, example 91 from intermediate 13) (23.0 g, 0.058mmol) and triethylamine (0.016 mL, 0.116 mmol) in CH₂Cl₂ (2 mL) wasadded 4-methylpiperazine-1-carbonyl chloride hydrochloride (11.6 g,0.058 mmol) and the mixture stirred at rt for 18 h. Diluted with CH₂Cl₂(10 mL) and washed with sat. NaHCO₃(aq) (2 mL). The organic layer wasfiltered through a phase separation tube and evaporated in vacuo.Purification was performed by reverse phase Gilson HPLC (Method A) andsubsequent neutralization of the combined fractions by elution throughan Isolute® SCX-2 cartridge, eluting with methanol, then with 2M ammoniain methanol. Basic fractions were concentrated in vacuo to give{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(4-methyl-piperazin-1-yl)-methanone(25 mg, 58% yield) as a yellow powder. ¹H NMR (400 MHz, CDCl₃, 298K) δppm 2.17-2.27 (m, 2H) 2.55 (s, 3H) 2.65-2.78 (m, 4H) 3.07 (t, 2H)3.45-3.73 (m, 9H) 3.86-3.95 (m, 1H) 4.02 (s, 3H) 4.13 (s, 2H) 5.66-5.73(m, 1H) 7.62 (d, 1H) 8.06 (d, 1H) 8.64 (s, 1H) LCMS: [M+H]+=522.3,Rt⁽¹⁾=1.21 min.

Example 86 was prepared using procedures analogous to those used inExample 85 using appropriate starting materials.

Example 86

Rt⁽¹⁾ (min.) 1.71 MS: [M + H]+ 509.2

Name:—{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-morpholin-4-yl-methanone

Purification method: Reverse phase HPLC Method A

Prepared using morpholine 4-carbonyl chloride

¹H NMR (400 MHz, MeOH-d4, 298K) δ ppm 2.21-2.31 (m, 2H) 3.03 (t, 2H)3.23-3.42 (m, 4H) 3.48-3.81 (m, 9H) 3.82-3.88 (m, 1H) 3.99 (s, 3H) 4.17(s, 2H) 5.72-5.77 (m, 1H) 7.79 (d, 1H) 8.13 (d, 1H) 8.59 (s, 1H)

Example 87:(4-Hydroxy-piperidin-1-yl)-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanone

To CH₂Cl₂ (5 mL) in a round bottomed flask was added phosgene (20%solution in toluene, 0.20 mL, 0.379 mmol) and the resulting solutioncooled to 5° C. under argon. A solution of6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(prepared using step 1, example 91 from intermediate 13) (50.0 g, 0.126mmol) and triethylamine (0.053 mL, 0.380 mmol) in CH₂Cl₂ (1.0 mL) wasadded and the mixture allowed to warm to room temperature with stirringunder argon over 1 h. Evaporated to dryness by bubbling a stream ofargon into the mixture to give a brown gum. Dissolved in CH₂Cl₂ (3 mL)to give(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonylchloride as a solution in CH₂Cl₂. LCMS: 458.4 [M+1]+, Rt⁽⁷⁾=1.38 min.This solution was used without further purification. 1.5 mL of thissolution was added to a solution of piperidin-4-ol (6.4 mg, 0.063 mmol)and triethylamine (0.053 mL, 0.380 mmol) in CH₂Cl₂ and the mixturestirred at room temperature under argon for 1 h. N,N-dimethylformamide(0.5 mL) was added and stirring continued for 2 h. Diluted with CH₂Cl₂(2 mL) and washed with sat. NaHCO₃(aq) (2 mL). The organic layer wasfiltered through a phase separation tube and evaporated in vacuo.Purification by reverse phase Gilson HPLC (Method A) and subsequentneutralization of the combined fractions by elution through an Isolute®SCX-2 cartridge, eluting with methanol, then with 2M ammonia inmethanol. Basic fractions were concentrated in vacuo to give(4-hydroxy-piperidin-1-yl)-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-methanoneas a pale yellow powder (22 mg, 64% yield). ¹H NMR (400 MHz, MeOH-d4,298K) δ ppm 1.35-1.57 (m, 2H) 1.80-.1.94 (m, 2H) 2.20-2.31 (m, 2H)2.94-3.09 (m, 4H) 3.45-3.87 (m, 10H) 3.98 (s, 3H) 4.17 (s, 2H) 5.70-5.76(m, 1H) 7.78 (d, 1H) 8.13 (d, 1H) 8.58 (s, 1H) LCMS: [M+H]+=523.2,Rt⁽¹⁾=1.58 min.

Example 88 was prepared using procedures analogous to those used inExample 87 using appropriate starting materials.

Example 88

Rt⁽¹⁾ (min.) 1.58 MS: [M + H]+ 497.2

Name:(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid (2-hydroxy-ethyl)-methyl-amide

Purification method: Reverse phase HPLC Method A

Prepared using 2-methylamino-ethanol

¹H NMR (400 MHz, MeOH-d4, 298K) δ ppm 2.20-2.32 (m, 2H) 2.97 (s, 3H)2.98-3.06 (t, 2H) 3.27-3.38 (m, 1H) 3.40-3.80 (m, 8H) 3.82-3.89 (m, 1H)3.98 (s, 3H) 4.18 (s, 2H) 5.71-5.76 (m, 1H) 7.78 (d, 1H) 8.14 (d, 1H)8.58 (s, 1H)

Example 89:1-(4-{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperazin-1-yl)-ethanone

To a solution of6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidine(prepared using step 1, example 91 from intermediate 13) (25 g, 0.063mmol) and triethylamine (0.013 mL, 0.095 mmol) in CH₂Cl₂ (2 mL) wasadded 3-(4-acetyl-piperazine-1-carbonyl)-1-methyl-3H-imidazol-3-iumiodide (Intermediate 6) (15 g, 0.063 mmol) and the mixture stirred atroom temperature under argon for 18 h.

Partitioned between CH₂Cl₂ (10 mL) and sat. NaHCO₃(aq) (2 mL) and theorganic layer was filtered through a phase separation tube andevaporated in vacuo. Purification by reverse phase Gilson HPLC (MethodA) and subsequent neutralization of the combined fractions by elutionthrough an Isolute® SCX-2 cartridge, eluting with methanol, then with 2Mammonia in methanol. Basic fractions were concentrated in vacuo to give1-(4-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperazin-1-yl)-ethanoneas a pale yellow powder (9 mg, 25% yield). ¹H NMR (400 MHz, MeOH-d4,298K) δ ppm 2.14 (s, 3H) 2.24-2.33 (m, 2H) 3.04 (t, 2H) 3.25-3.91 (m,13H) 3.99 (s, 3H) 4.18 (s, 2H) 5.74-5.78 (m, 1H) 7.79 (d, 1H) 8.14 (d,1H) 8.60 (s, 1H) LCMS: [M+H]+=550.2, Rt⁽¹⁾=1.58 min.

Example 90:(S)-3-[6-(5-cyano-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid methyl ester

To a solution of2-methoxy-5-[4-((S)-pyrrolidin-3-yloxy)-7,8-dihydro-5H-pyrido[4,3-d]pyrimidin-6-yl]-nicotinonitrile(prepared using intermediate 11 and method Ib, process step 2,example 1) (25.0 g, 0.071 mmol) and triethylamine (0.04 mL, 0.29 mmol)in CH₂Cl₂ (2 mL) was added methyl carbonochloridate (0.006 mL, 0.078mmol) and the mixture stirred at room temperature for 18 h. Diluted withCH₂Cl₂ (2 mL) and washed with sat. NaHCO₃(aq) (1 mL). The organic layerwas filtered through a phase separation tube and evaporated in vacuo.Purification was performed by reverse phase Gilson HPLC (Method A) andsubsequent neutralization of the combined fractions by elution throughan Isolute® SCX-2 cartridge, eluting with methanol, then with 2M ammoniain methanol. Basic fractions were concentrated in vacuo gave(S)-3-[6-(5-cyano-6-methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid methyl ester (10 mg, 35% yield) as a yellow powder. ¹H NMR (400MHz, DMSO-d6, 298K) δ ppm 2.09-2.31 (m, 2H) 2.91 (t, 2H) 3.45-3.75 (m,9H) 3.93 (s, 3H) 4.17 (s, 2H) 5.58-5.65 (m, 1H) 8.09 (d, 1H) 8.27 (d,1H) 8.61 (s, 1H) LCMS: [M+H]+=411.1, Rt⁽¹⁼1.58 min.

Example 91:{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanone

Step 2

To the oxazole-4-carboxylic acid (27 g, 0.24 mmol)) and HBTU (89 g, 0.24mmol) in DMF (1 mL) was added N,N-diisopropylethylamine (0.08 mL, 0.45mmol). The mixture was stirred for 20 min and then6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidinedihydrochloride (100 g, 0.214 mmol) and additionalN,N-diisopropylethylamine (0.08 mL, 0.45 mmol) were added. The mixturewas allowed to stir at room temperature for 30 min. Purified by reversephase Gilson HPLC (Method A) and subsequent neutralization of thecombined fractions over PL-HCO₃ MP to give{(S)-3-[6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-4-yl-methanoneas a yellow solid (38 mg, 36% yield) ¹H NMR (400 MHz, DMSO-d6, 298K) δppm 2.11-2.39 (m, 2H) 2.80-3.01 (m, 2H) 3.22-4.29 (m, 11H) 5.59-5.80 (m,1H) 7.72-7.94 (m, 1H) 8.10-8.29 (m, 1H) 8.41-8.55 (m, 1H) 8.57-8.77 (m,2H) LCMS: [M+H]+=491.1, Rt⁽¹⁾=1.69 min.

6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidinedihydrochloride

Step 1

(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carboxylicacid tert-butyl ester (1.0 g, 1.69 mmol) in CH₂Cl₂ (5 mL) was added 2Manhydrous HCl in diethyl ether (25.3 mL, 50.5 mmol) and the mixturestirred at rt for 3 h. The resulting precipitate was filtered and washedwith diethyl ether to give6-(6-methoxy-5-trifluoromethyl-pyridin-3-yl)-4-((S)-pyrrolidin-3-yloxy)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidinedihydrochloride as a yellow solid (1.01 g, 128% yield). [M+H]+=396.0,Rt⁽⁴⁾=0.71 min. The free base can be generated by partitioning thedihydrochloride salt between dichloromethane and 1N sodium hydroxidesolution(aq), separating the organic phase and evaporating in vacuo.[M+H]+=396.0, Rt⁽⁴⁾=0.71 min.

Example 92 was prepared using procedures analogous to those used inExample 91 using appropriate starting materials.

Example 92

Rt⁽¹⁾ (min.) 1.58 MS: [M + H]+ 549.2

Name:1-(4-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidine-1-carbonyl}-piperidin-1-yl)-ethanone

Purification method: Reverse phase method A

Prepared using 1-acetyl-piperidine-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.21-1.77 (m, 4H) 1.92-2.02 (m,3H) 2.08-2.36 (m, 2H) 2.42-2.80 (m, 2H) 2.88-2.98 (m, 2H) 3.00-3.18 (m,1H) 3.39-4.24 (m, 13H) 5.60-5.74 (m, 1H) 7.80-7.87 (m, 1H) 8.15-8.22 (m,1H) 8.59-8.65 (m, 1H)

Example 93

Rt⁽¹⁾ (min.) 1.37 MS: [M + H]+ 504.1

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-(3-methyl-3H-imidazol-4-yl)-methanone

Purification method: Reverse phase method A

Prepared using 3-methyl-3H-imidazole-4-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ pm 2.14-2.38 (m, 2H) 2.78-3.08 (m, 2H)3.44-4.04 (m, 12H) 4.08-4.27 (m, 2H) 5.66-5.73 (m, 1H) 7.32-7.57 (m, 1H)7.70-7.97 (m, 2H) 8.13-8.28 (m, 1H) 8.56-8.69 (m, 1H)

Example 94

Rt⁽¹⁾ (min.) 0.95 MS: [M + H]+ 490.9

Name:{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy]-pyrrolidin-1-yl}-oxazol-5-yl-methanone

Purification method: Reverse phase method A

Prepared using oxazole-5-carboxylic acid

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 2.28-2.43 (m, 2H) 2.86-3.00 (m,2H) 3.39-4.27 (m, 11H) 5.60-5.85 (m, 1H) 7.72-7.91 (m, 2H) 8.15-8.30 (m,1H) 8.53-8.68 (m, 2H)

Example 95 was prepared using procedures analogous to those used inExample 1, method 1a using appropriate starting materials according toscheme 8.

Example Structure Rt⁽³⁾ (min.) MS: [M + H]+ 95

1.39( 440.1

Name:{(S)-3-[6-(6-Methoxy-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-yloxy)pyrrolidin-1-yl}-(tetrahydro-pyran-4-yl)-methanone

Purification method: Flash-chromatography on silica gel with CH₂Cl₂/MeOH

Prepared using 1-benzyl-1-methyl-4-oxo-piperidinium iodide (Ref:Tortolani, R.; Org. Lett., Vol. 1, No 8, 1999) and 2-methoxypyridine

¹H NMR (400 MHz, DMSO-d6, 298K) δ ppm 1.48-1.65 (m, 4H) 2.05-2.30 (m,2H) 2.59-2.78 (m, 1H) 2.85-2.93 (m, 2H) 3.25-4.11 (m, 15H) 5.59-5.73 (m,1H) 6.73-6.79 (m, 1H) 7.53-7.59 (m, 1H) 7.86-7.89 (m, 1H) 8.58-8.64 (m,1H).

The pharmaceutical composition or combination of the present inventioncan be in unit dosage of about 1-2000 mg of active ingredient(s) for asubject of about 50-70 kg, or about 1-500 mg or about 1-250 mg or about1-150 mg or about 0.5-100 mg, or about 1-50 mg of active ingredients.The therapeutically effective dosage of a compound, the pharmaceuticalcomposition, or the combinations thereof, is dependent on the species ofthe subject, the body weight, age and individual condition, the disorderor disease or the severity thereof being treated. A physician, clinicianor veterinarian of ordinary skill can readily determine the effectiveamount of each of the active ingredients necessary to prevent, treat orinhibit the progress of the disorder or disease.

The above-cited dosage properties are demonstrable in vitro and in vivotests using advantageously mammals, e.g., mice, rats, dogs, monkeys orisolated organs, tissues and preparations thereof. The compounds of thepresent invention can be applied in vitro in the form of solutions,e.g., aqueous solutions, and in vivo either enterally, parenterally,advantageously intravenously, e.g., as a suspension or in aqueoussolution. The dosage in vitro may range between about 10⁻³ molar and10⁻⁹ molar concentrations. A therapeutically effective amount in vivomay range depending on the route of administration, between about0.1-500 mg/kg, or between about 1-100 mg/kg.

Biological Evaluation

The activity of a compound according to the present invention can beassessed by the following in vitro & in vivo methods.

Biological Assays

1 Determination of Enzymatic PI3K Alpha and PI3K Delta IsoformInhibition

1.1 Test of Lipid Kinase Activity

The efficacy of the compounds of examples 1-117 as PI3 kinase inhibitorscan be demonstrated as follows:

The kinase reaction is performed in a final volume of 50 μl per well ofa half area COSTAR, 96 well plate. The final concentrations of ATP andphosphatidyl inositol in the assay are 5 μM and 6 μg/mL, respectively.The reaction is started by the addition of PI3 kinase, e.g. PI3 kinaseδ.

p110δ. The components of the assay are added per well as follows:

-   -   10 μl test compound in 5% DMSO per well in columns 2-1.    -   Total activity is determined by addition 10 μl of 5% vol/vol        DMSO in the first 4 wells of column 1 and the last 4 wells of        column 12.    -   The background is determined by addition of 10 μM control        compound to the last 4 wells of column 1 and the first 4 wells        of column 12.    -   2 mL ‘Assay mix’ are prepared per plate:        -   1.912 mL of HEPES assay buffer        -   8.33 μl of 3 mM stock of ATP giving a final concentration of            5 μM per well        -   1 μl of [³³P]ATP on the activity date giving 0.05 μCi per            well        -   1 μl of 1 mg/mL PI stock giving a final concentration of 6            μg/mL per well        -   1 μl of 1 M stock MgCl₂ giving a final concentration of 1 mM            per well    -   20 μl of the assay mix are added per well.    -   2 mL ‘Enzyme mix’ are prepared per plate (x*μl PI3 kinase p110β        in 2 mL of kinase buffer). The ‘Enzyme mix’ is kept on ice        during addition to the assay plates.    -   20 μl ‘Enzyme mix’ are added/well to start the reaction.    -   The plate is then incubated at room temperature for 90 minutes.    -   The reaction is terminated by the addition of 50 μl WGA-SPA bead        (wheat germ agglutinin-coated Scintillation Proximity Assay        beads) suspension per well.    -   The assay plate is sealed using TopSeal-S (heat seal for        polystyrene microplates, PerkinElmer LAS [Deutschland] GmbH,        Rodgau, Germany) and incubated at room temperature for at least        60 minutes.    -   The assay plate is then centrifuged at 1500 rpm for 2 minutes        using the Jouan bench top centrifuge (Jouan Inc., Nantes,        France).    -   The assay plate is counted using a Packard TopCount, each well        being counted for 20 seconds.    -   The volume of enzyme is dependent on the enzymatic activity of        the batch in use.

In a more preferred assay, the kinase reaction is performed in a finalvolume of 10 μl per well of a low volume non-binding CORNING, 384 wellblack plate (Cat. No. #3676). The final concentrations of ATP andphosphatidyl inositol (PI) in the assay are 1 μM and 10 μg/mL,respectively. The reaction is started by the addition of ATP.

The components of the assay are added per well as follows:

50 nl test compounds in 90% DMSO per well, in columns 1-20, 8concentrations (1/3 and 1/3.33 serial dilution step) in single.

-   -   Low control: 50 nl of 90% DMSO in half the wells of columns        23-24 (0.45% in final).    -   High control: 50 nl of reference compound (e.g. compound of        Example 7 in WO 2006/122806) in the other half of columns 23-24        (2.5 μM in final).    -   Standard: 50 nl of reference compound as just mentioned diluted        as the test compounds in columns 21-22.    -   20 mL ‘buffer’ are prepared per assay:        -   200 μl of 1M TRIS HCl pH7.5 (10 mM in final)        -   60 μl of 1M MgCl₂ (3 mM in final)        -   500 μl of 2M NaCl (50 mM in final)        -   100 μl of 10% CHAPS (0.05% in final)        -   200 μl of 100 mM DTT (1 mM in final)        -   18.94 mL of nanopure water    -   10 mL ‘PI’ are prepared per assay:        -   200 μl of 1 mg/mL 1-alpha-Phosphatidylinositol (Liver            Bovine, Avanti Polar Lipids Cat. No. 840042C MW=909.12)            prepared in 3% OctylGlucoside (10 μg/mL in final)        -   9.8 mL of ‘buffer’    -   10 mL ‘ATP’ are prepared per assay:        -   6.7 μl of 3 mM stock of ATP giving a final concentration of            1 μM per well        -   10 mL of ‘buffer’    -   2.5 mL of each PI3K construct are prepared per assay in ‘PI’        with the following final concentration:        -   10 nM PI3K alfa EMV B1075        -   25 nM beta EMV BV949        -   10 nM delta EMV BV1060        -   150 nM gamma EMV BV950    -   5 μl of ‘PI/PI3K’ are added per well.    -   5 μl ‘ATP’ are added per well to start the reaction.    -   The plates are then incubated at room temperature for 60 minutes        (alfa, beta, delta) or 120 minutes (gamma).    -   The reaction is terminated by the addition of 10 μl Kinase-Glo        (Promega Cat. No. #6714).    -   The assay plates are read after 10 minutes in Synergy 2 reader        (BioTek, Vermont USA) with an integration time of 100        milliseconds and sensitivity set to 191.    -   Output: The High control is around 60′000 counts and the Low        control is 30′000 or lower    -   This luminescence assay gives a useful Z′ ratio between 0.4 and        0.7

The Z′ value is a universal measurement of the robustness of an assay. AZ′ between 0.5 and 1.0 is considered an excellent assay.

For this assay, the PI3K constructs mentioned are prepared as follows:

1.2 Generation of Gene Constructs

Two different constructs, BV 1052 and BV 1075, are used to generate thePI3 Kinase α proteins for compound screening.

PI3Kα BV-1052 p85(iSH2)-Gly Linker-p110a (D20aa)-C-Term his Tag

PCR products for the inter SH2 domain (iSH2) of the p85 subunit and forthe p110-a subunit (with a deletion of the first 20 amino acids) aregenerated and fused by overlapping PCR. The iSH2 PCR product isgenerated from first strand cDNA using initially primers gwG130-p01(5′-CGAGAATATGATAGATTATATGAAGAAT-3′) (SEQ ID NO: 1) and gwG130-p02(5′-TGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′) (SEQ ID NO: 2). Subsequently ina secondary PCR reaction, Gateway (Invitrogen AG, Basel, Switzerland)recombination AttB1 sites and linker sequences are added at the 5′endand 3′end of the p85 iSH2 fragment respectively, using primersgwG130-p03(5′-GGGACAAGTTTGTACAAAAAAGCAGGCTACGAAGGAGATATACATAT-GCGAGAATATGATAGATTATATGAAGAAT-3′)(SEQ ID NO: 3) and gwG152-p04(5′-TACCATAATTCCACCACCACCACCGGAAATTCCCCCTGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′)(SEQ ID NO: 4).

The p110-a fragment is also generated from first strand cDNA, initiallyusing primers gwG152-p01 (5′-CTAGTGGAATGTTTACTACCAAATGG-3′) (SEQ ID NO:5) and gwG152-p02 (5′-GTTCAATG-CATGCTGTTTAATTGTGT-3′) (SEQ ID NO: 6).

In a subsequent PCR reaction, linker sequence and a Histidine tag areadded at the 5′end and 3′end of the p110-a fragment respectively, usingprimers gw152-p03(5′-GGGGGAATTTCCGGTGGTGGTGGTGGAATTATGGTAC-TAGTGGAATGTTTACTACC-AAATGGA-3′)(SEQ ID NO: 7) and gwG152-p06(5′-AGCTCCGTGATGGTGATGGTGATGTGCTCCGTTCAATG-CATGCTGTTTAATTGTGT-3′) (SEQID NO: 8).

The p85-iSH2/p110-a fusion protein is assembled in a third PCR reactionby the overlapping linkers at the 3′end of the iSH2 fragment and the5′end of the p110-a fragment, using the above mentioned gwG130-p03primer and a primer containing an overlapping Histidine tag and theAttB2 recombination sequences(5′-GGGACCACTTTGTACAAGAAAGCTGGGTTTAAGCTCCGTGATGGTGATGGTGAT-GTGCTCC-3′)(SEQ ID NO: 9).

This final product is recombined in a (Invitrogen) OR reaction into thedonor vector pDONR201 to generate the ORF318 entry clone. This clone isverified by sequencing and used in a Gateway LR reaction to transfer theinsert into the Gateway adapted pBlueBac4.5 (Invitrogen) vector forgeneration of the baculovirus expression vector LR410.

PI3Kα BV-1075 p85(iSH2)-12 XGly Linker-p110α (D20aa)-C-Term his Tag

The construct for Baculovirus BV-1075 is generated by a three-partligation comprised of a p85 fragment and a p110-a fragment cloned intovector pBlueBac4.5. The p85 fragment is derived from plasmid p1661-2digested with Nhe/Spe. The p110-a fragment derived from LR410 (seeabove) as a SpeI/HindIII fragment. The cloning vector pBlueBac4.5(Invitrogen) is digested with Nhe/HindIII. This results in the constructPED 153.8

The p85 component (iSH2) is generated by PCR using ORF 318 (describedabove) as a template and one forward primer KAC1028(5′-GCTAGCATGCGAGAATATGATAGATTATATGAAGAATATACC) (SEQ ID NO: 10) and tworeverse primers, KAC1029(5′-GCCTCCACCACCTCCGCCTGGTTTAATGCTGTTCATACGTTTGTC) (SEQ ID NO: 11) andKAC1039 (5′-TACTAGTCCGCCTCCACCACCTCCGCCTCCACCACCTCCGCC) (SEQ ID NO: 12).

The two reverse primers overlap and incorporate the 12× Gly linker andthe N-terminal sequence of the p110a gene to the SpeI site. The 12× Glylinker replaces the linker in the BV1052 construct. The PCR fragment iscloned into pCR2.1 TOPO (Invitrogen). Of the resulting clones, p1661-2is determined to be correct. This plasmid is digested with Nhe and SpeIand the resulting fragment is gel-isolated and purified for sub-cloning.

The p110-a cloning fragment is generated by enzymatic digest of cloneLR410 (see above) with Spe I and HindIII. The SpeI site is in the codingregion of the p110α gene. The resulting fragment is gel-isolated andpurified for sub-cloning.

The cloning vector, pBlueBac4.5 (Invitrogen) is prepared by enzymaticdigestion with Nhe and HindIII. The cut vector is purified with Qiagen(Quiagen N.V, Venlo, Netherlands) column and then dephosphorylated withCalf Intestine alkaline phosphatase (CIP) (New England BioLabs, Ipswich,Mass.). After completion of the CIP reaction the cut vector is againcolumn purified to generate the final vector. A 3 part ligation isperformed using Roche Rapid ligase and the vendor specifications.

PI3Kβ BV-949 p85(iSH2)-Gly Linker-p110b(Full-Length)-C-Term his Tag

PCR products for the inter SH2 domain (iSH2) of the p85 subunit and forthe full-length p110-b subunit are generated and fused by overlappingPCR.

The iSH2 PCR product is generated from first strand cDNA initially usingprimers gwG130-p01 (5′-CGAGAATATGATAGATTATATGAAGAAT-3′) (SEQ ID NO: 1)and gwG130-p02 (5′-TGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′) (SEQ ID NO: 2).Subsequently, in a secondary PCR reaction Gateway (Invitrogen)recombination AttB1 sites and linker sequences are added at the 5′endand 3′end of the p85 iSH2 fragment respectively, using primersgwG130-p03(5′-GGGACAAGTTTGTACAAAAAAGCAGGCTACGAAGGAGATA-TACATATGCGAGAATATGATAGATTATATGAAGAAT-3′)(SEQ ID NO: 3) and gwG130-p05(5′-ACTGAAGCATCCTCCTCCTCCTCCTCCTGGTTTAAT-GCTGTTCATACGTTTGTC-3′) (SEQ IDNO: 13).

The p110-b fragment is also generated from first strand cDNA initiallyusing primers gwG130-p04(5′-ATTAAACCAGGAGGAGGAGGAGGAGGATGCTTCAGTTTCATAATGCC-TCCTGCT-3′) (SEQ IDNO: 4) which contains linker sequences and the 5′end of p110-b andgwG130-p06(5′-AGCTCCGTGATGGTGATGGTGATGTGCTCCAGATCTGTAGTCTTT-CCGAACTGTGTG-3′) (SEQID NO: 14) which contains sequences of the 3′end of p110-b fused to aHistidine tag.

The p85-iSH2/p110-b fusion protein is assembled by an overlapping PCR areaction of the linkers at the 3′end of the iSH2 fragment and the 5′endof the p110-b fragment, using the above mentioned gwG130-p03 primer anda primer containing an overlapping Histidine tag and the AttB2recombination sequences(5′-GGGACCACTTTGTACAAGAAAGCTGGGTTT-AAGCTCCGTGATGGTGATGGTGATGTGCTCC-3′)(SEQ ID NO: 15).

This final product is recombined in a Gateway (Invitrogen) OR reactioninto the donor vector pDONR201 to generate the ORF253 entry clone. Thisclone is verified by sequencing and used in a Gateway LR reaction totransfer the insert into the Gateway adapted pBlueBac4.5 (Invitrogen)vector for generation of the baculovirus expression vector LR280.

PI3Kδ BV-1060 p85(iSH2)-Gly Linker-p110d(Full-Length)-C-Term his Tag

PCR products for the inter SH2 domain (iSH2) of the p85 subunit and forthe full-length p110-d subunit are generated and fused by overlappingPCR.

The iSH2 PCR product is generated from first strand cDNA using initiallyprimers gwG130-p01 (5′-CGAGAATATGATAGATTATATGAAGAAT-3′) (SEQ ID NO: 1)and gwG130-p02 (5′-TGGTTT-AATGCTGTTCATACGTTTGTCAAT-3′) (SEQ ID NO: 2).Subsequently, in a secondary PCR reaction Gateway (Invitrogen)recombination AttB1 sites and linker sequences are added at the 5′endand 3′end of the p85 iSH2 fragment respectively, using primersgwG130-p03(5′-GGGACAAGTTTGTACAAAAAAGCAGGCTACGAAGGAGATATACAT-ATGCGAGAATATGATAGATTATATGAAGAAT-3′)(SEQ ID NO: 3) and gwG154-p04(5′-TCCTCCTCCTCCTCCTCCTGGTTTAATGCTGTTCATACGTTTGTC-3′) (SEQ ID NO: 16).

The p110-a fragment is also generated from first strand cDNA usinginitially primers gwG154-p01 (5′-ATGCCCCCTGGGGTGGACTGCCCCAT-3′) (SEQ IDNO: 17) and gwG154-p02 (5′-CTACTG-CCTGTTGTCTTTGGACACGT-3′) (SEQ ID NO:18).

In a subsequent PCR reaction linker sequences and a Histidine tag isadded at the 5′end and 3′end of the p110-d fragment respectively, usingprimers gw154-p03(5′-ATTAAACCAGGAGGAGGAGGAGGAGGACCCCCTGGGGTGGAC-TGCCCCATGGA-3′) (SEQ IDNO: 19) and gwG154-p06(5′-AGCTCCGTGATGGTGAT-GGTGATGTGCT-CCCTGCCTGTTGTCTTTGGACACGTTGT-3′) (SEQID NO: 20).

The p85-iSH2/p110-d fusion protein is assembled in a third PCR reactionby the overlapping linkers at the 3′end of the iSH2 fragment and the5′end of the p110-d fragment, using the above mentioned gwG130-p03primer and a primer containing an overlapping Histidine tag and theGateway (Invitrogen) AttB2 recombination sequences(5′-GGGACCACTTTGTA-CAAGAAAGCTGGGTTT-AAGCTCCGTGATGGTGATGGTGATGTGCTCC-3′)(SEQ ID NO: 21).

This final product is recombined in a Gateway (Invitrogen) OR reactioninto the donor vector pDONR201 to generate the ORF319 entry clone. Thisclone is verified by sequencing and used in a Gateway LR reaction totransfer the insert into the Gateway adapted pBlueBac4.5 (Invitrogen)vector for generation of the baculovirus expression vector LR415.

PI3Kγ BV-950 p110g(D144aa)-C-Term his Tag

This construct is obtained from Roger Williams lab, MRC Laboratory ofMolecular Biology, Cambridge, UK (November, 2003). Description of theconstruct in: Pacold M. E. et al. (2000) Cell 103, 931-943.

1.3 Protein Expression and Purification

Methods to generate recombinant baculovirus and protein for PI3Kisoforms:

The pBlue-Bac4.5 (for a, b, and d isoforms) or pVL1393 (for g) plasmidscontaining the different PI3 kinase genes are co-transfected withBaculoGold WT genomic DNA (BD Biosciences, Franklin Lakes, N.J., USA)using methods recommended by the vendor. Subsequently, the recombinantbaculovirus obtained from the transfection is plaque-purified on Sf9insect cells to yield several isolates expressing recombinant protein.Positive clones are selected by anti-HIS or anti-isoform antibodywestern. For PI3K alpha and delta isoforms, a secondaryplaque-purification is performed on the first clonal virus stocks ofPI3K. Amplification of all baculovirus isolates is performed at lowmultiplicity of infection (moi) to generate high-titer, low passagestock for protein production. The baculoviruses are designated BV1052(α) and BV1075 (α), BV949 (β), BV1060 (δ) and BV950 (γ).

Protein production involves infection (passage 3 or lower) of suspendedTn5 (Trichoplusia ni) or TiniPro (Expression Systems, LLC, Woodland,Calif., USA) cells in protein-free media at moi of 2-10 for 39-48 hoursin 2 l glass Erlenmyer flasks (110 rpm) or wave-bioreactors (22-25 rpm).Initially, 10 l working volume wave-bioreactors are seeded at a densityof 3e5 cells/mL at half capacity (5 L). The reactor is rocked at 15 rpmduring the cell growth phase for 72 hours, supplemented with 5% oxygenmixed with air (0.2 l per minute). Immediately prior to infection, thewave-reactor cultures are analyzed for density, viability and diluted toapproximately 1.5e6 cell/mL. 100-500 mL of high titer, low passage virusis added following 2-4 hours of additional culture. Oxygen is increasedto 35% for the 39-48 hour infection period and rocking platform rpmincreased to 25. During infection, cells are monitored by Vicellviability analyzer (Beckman Coulter, Inc, Fullerton, Calif., USA)bioprocess for viability, diameter and density. Nova Bioanalyzer (NOVABiomedical Corp., Waltham, Mass., USA) readings of various parametersand metabolites (pH, O₂ saturation, glucose, etc.) are taken every 12-18hours until harvest. The wave-bioreactor cells are collected within 40hours post infection. Cells are collected by centrifugation (4 degreesC. at 1500 rpm), and subsequently maintained on ice during pooling ofpellets for lysis and purification. Pellet pools are made with smallamounts of cold, un-supplemented Grace's media (w/o proteaseinhibitors).

PI3K Alpha Purification Protocol for HTS (BV1052)

PI3K alpha is purified in three chromatographic steps: immobilized metalaffinity chromatography on a Ni Sepharose resin (GE Healthcare,belonging to General Electric Company, Fairfield, Conn., USA), gelfiltration utilizing a Superdex 200 26/60 column (GE Healthcare), andfinally a cation exchange step on a SP-XL column (GE Healthcare). Allbuffers are chilled to 4° C. and lysis is performed chilled on ice.Column fractionation is performed rapidly at room temperature.

Typically frozen insect cells are lysed in a hypertonic lysis buffer andapplied to a prepared IMAC column. The resin is washed with 3-5 columnvolumes of lysis buffer, followed by 3-5 column volumes wash buffercontaining 45 mM imidazole, and the target protein is then eluted with abuffer containing 250 mM imidazole. Fractions are analyzed by Coomassiestained SDS-PAGE gels, and fractions containing target protein arepooled and applied to a prepared GFC column. Fractions from the GFCcolumn are analyzed by Coomassie stained SDS-PAGE gels, and fractionscontaining target protein are pooled. The pool from the GFC column isdiluted into a low salt buffer and applied to a prepared SP-XL column.The column is washed with low salt buffer until a stable A280 baselineabsorbance is achieved, and eluted using a 20 column volume gradientfrom 0 mM NaCl to 500 mM NaCl. Again, fractions from the SP-XL columnare analyzed by Coomassie stained SDS-PAGE gels, and fractionscontaining the target protein are pooled. The final pool is dialyzedinto a storage buffer containing 50% glycerol and stored at −20° C. Thefinal pool is assayed for activity in a phosphoinosititol kinase assay.

PI3K Beta Purification Protocol for HTS (BV949)

PI3K beta is purified in two chromatographic steps: immobilized metalaffinity chromatography (IMAC) on a Ni Sepharose resin (GE Healthcare)and gel filtration (GFC) utilizing a Superdex 200 26/60 column (GEHealthcare). All buffers are chilled to 4° C. and lysis is performedchilled on ice. Column fractionation is performed rapidly at roomtemperature.

Typically frozen insect cells are lysed in a hypertonic lysis buffer andapplied to a prepared IMAC column. The resin is washed with 3-5 columnvolumes of lysis buffer, followed by 3-5 column volumes wash buffercontaining 45 mM imidazole, and the target protein is then eluted with abuffer containing 250 mM imidazole. Fractions are analyzed by Coomassiestained SDS-PAGE gels, and fractions containing target protein arepooled and applied to a prepared GFC column. Fractions from the GFCcolumn are analyzed by Coomassie stained SDS-PAGE gels, and fractionscontaining target protein are pooled. The final pool is dialyzed into astorage buffer containing 50% glycerol and stored at −20° C. The finalpool is assayed for activity in the phosphoinostitol kinase assay.

PI3K Gamma Purification Protocol for HTS (BV950)

PI3K gamma is purified in two chromatographic steps: immobilized metalaffinity chromatography (IMAC) on a Ni Sepharose resin (GE Healthcare)and gel filtration (GFC) utilizing a Superdex 200 26/60 column (GEHealthcare). All buffers are chilled to 4° C. and lysis is performedchilled on ice. Column fractionation is performed rapidly at roomtemperature. Typically frozen insect cells are lysed in a hypertoniclysis buffer and applied to a prepared IMAC column. The resin is washedwith 3-5 column volumes of lysis buffer, followed by 3-5 column volumeswash buffer containing 45 mM imidazole, and the target protein is theneluted with a buffer containing 250 mM imidazole. Fractions are analyzedby Coomassie stained SDS-PAGE gels, and fractions containing targetprotein are pooled and applied to a prepared GFC column. Fractions fromthe GFC column are analyzed by Coomassie stained SDS-PAGE gels, andfractions containing target protein are pooled. The final pool isdialyzed into a storage buffer containing 50% glycerol and stored at−20° C. The final pool is assayed for activity in the phosphoinostitolkinase assay.

PI3K Delta Purification Protocol for HTS (BV1060)

PI3K delta is purified in three chromatographic steps: immobilized metalaffinity chromatography on a Ni Sepharose resin (GE Healthcare), gelfiltration utilizing a Superdex 200 26/60 column (GE Healthcare), andfinally a anion exchange step on a Q-HP column (GE Healthcare). Allbuffers are chilled to 4° C. and lysis is performed chilled on ice.Column fractionation is performed rapidly at room temperature. Typicallyfrozen insect cells are lysed in a hypertonic lysis buffer and appliedto a prepared IMAC column. The resin is washed with 3-5 column volumesof lysis buffer, followed by 3-5 column volumes wash buffer containing45 mM imidazole, and the target protein is then eluted with a buffercontaining 250 mM imidazole. Fractions are analyzed by Coomassie stainedSDS-PAGE gels, and fractions containing the target protein are pooledand applied to a prepared GFC column. Fractions from the GFC column areanalyzed by Coomassie stained SDS-PAGE gels, and fractions containingthe target protein are pooled. The pool from the GFC column is dilutedinto a low salt buffer and applied to a prepared Q-HP column. The columnis washed with low salt buffer until a stable A280 baseline absorbanceis achieved, and eluted using a 20 column volume gradient from 0 mM NaClto 500 mM NaCl. Again, fractions from the Q-HP column are analyzed byCoomassie stained SDS-PAGE gels, and fractions containing the targetprotein are pooled. The final pool is dialyzed into a storage buffercontaining 50% glycerol and stored at −20° C. The final pool is assayedfor activity in the phosphoinostitol kinase assay.

IC₅₀ is determined by a four parameter curve fitting routine that comesalong with “excel fit”. A four parameter logistic equation is used tocalculate IC₅₀ values (IDBS XLfit) of the percentage inhibition of eachcompound at 8 concentrations (usually 10, 3.0, 1.0, 0.3, 0.1, 0.030,0.010 and 0.003 μM). Alternatively, IC₅₀ values are calculated usingidbsXLfit model 204, which is a 4 parameter logistic model.

Yet alternatively, for an ATP depletion assay, compounds of the formulaI to be tested are dissolved in DMSO and directly distributed into awhite 384-well plate at 0.5 μl per well. To start the reaction, 10 μl of10 nM PI3 kinase and 5 μg/mL 1-alpha-phosphatidylinositol (PI) are addedinto each well followed by 10 μl of 2 μM ATP. The reaction is performeduntil approx 50% of the ATP is depleted, and then stopped by theaddition of 20 μl of Kinase-Glo solution (Promega Corp., Madison, Wis.,USA). The stopped reaction is incubated for 5 minutes and the remainingATP is then detected via luminescence. IC₅₀ values are then determined.

Some of the compounds of examples 1-49 and 51-95 show a certain level ofselectivity against the different paralogs PI3K α, β, γ and δ.

Suitably, the compounds of examples 1-49 and 51-95 show a certain levelof selectivity for the isoform PI3Kδ, e.g. as indicated in in vitro andin vivo tests against the different paralogs PI3K α and

β.

The range of activity, expressed as IC₅₀, in these assays, is preferablybetween 1 nM and 5000 nM, more preferably between 1 nM and about 1000nM.

2. Cellular Assays

2.1 Phosphoinositide-3 Kinase (PI3K)-Mediated Akt 1/2 (S473)Phosphorylation in Rat-1 Cells

Rat-1 cells stably overexpressing a myristoylated form of the catalyticsubunit of human phosphoinositide-3 kinase (PI3K) alpha, beta or deltawere plated in 384-well plates at a density of 7500 (PI3K alpha), 6200(PI3K beta), or 4000 (PI3K delta) cells in 30 ul complete growth medium(Dulbecco's modified Eagle's medium (DMEM high glucose) supplementedwith 10% (v/v) fetal bovine serum, 1% (v/v) MEM non essential aminoacids, 10 mM HEPES, 2 mM L-glutamine, 10 μg/mL puromycin and 1% (v/v)Penicillin/Streptomycin) and were incubated at 37% C/5% CO₂/95% humidityfor 24 h. Compounds were diluted in 384-well compound plates to obtain8-point serial dilutions for 40 test compounds in 90% DMSO, as well as 4reference compounds plus 16 high controls and 16 low (inhibited)controls.

Predilution plates were prepared by dispensing pipetting 250 nl ofcompound solutions into 384-well polypropylen plates using a Hummingwellnanoliter dispensor. Compounds were prediluted by the addition of 49.75ul complete growth medium. 10 ul of prediluted compound solution weretransferred to the cell plate using a 384-well pipettor, resulting in afinal DMSO concentration of 0.11%. Cells were incubated for 1 h at 37%C/5% CO₂/95% humidity. The supernatant was removed, the cells were lysedin 20 ul of lysis buffer for AlphaScreen® SureFire® detection.

For detection of p-AKT(Ser473), the SureFire® p-Akt 1/2 (Ser473) AssayKit (PerkinElmer, U.S.A) was used. 5 ul of cell lysate was transferredto 384-well low volume Proxiplates for detection using a 384-wellpipettor. Addition of AlphaScreen® SureFire® reagents was done accordingto the manufacturer's protocol. First, 5 ul of reaction buffer plusactivation buffer mix containing AlphaScreen® acceptor beads was added,the plate was sealed, and incubated on a plate shaker for 2 hours atroom temperature. Second, 2 ul of dilution buffer containingAlphaScreen® donor beads was added, and the plate was incubated on plateshaker as above for a further 2 hours. The plate was read on anAlphaScreen® compatible plate reader, using standard AlphaScreen®settings.

2.2 Determination of Murine B Cell Activation

PI3Kδ has been recognized to modulate B cell function when cells arestimulated through the B cell receptor (BCR) (Okkenhaug et al. Science297:1031 (2002). For assessing the inhibitory property of compounds on Bcell activation, the upregulation of activation markers CD86 and CD69 onmurine B cells derived from mouse spleen antibody is measured afterstimulation with anti-IgM. CD69 is a well known activation marker for Band T cells (Sancho et al. Trends Immunol. 26:136 (2005). CD86 (alsoknown as B7-2) is primarily expressed on antigen-presenting cells,including B cells. Resting B cells express CD86 at low levels, butupregulate it following stimulation of e.g. the BCR or IL-4 receptor.CD86 on a B cell interacts with CD28 on T cells. This interaction isrequired for optimal T cell activation and for the generation of anoptimal IgG1 response (Carreno et al. Annu Rev Immunol. 20:29 (2002)).

Spleens from Balb/c mice were collected, splenocytes were isolated andwashed twice with RPMI containing 10% foetal bovine serum (FBS), 10 mMHEPES, 100 Units/mL penicilline/streptomycine. RPMI supplemented in thisway is subsequently referred to as medium. The cells were adjusted to2.5×10⁶ cells/mL in medium and 200 μl cell suspension (5×10 cells) wereadded to the appropriate wells of 96 well plates.

Then the cells were stimulated by adding 50 μl anti-IgM mAb in medium(final concentration: 30 pg/mL). After incubation for 24 hours at 37°C., the cells were stained with the following antibody cocktails:anti-mouse CD86-FITC, anti-mouse CD69-PerCP-Cy5.5, anti-mouse CD19-PerCPfor the assessment of B cells, and anti-mouse CD3-FITC, anti-mouseCD69-PE for the assessment of T cells (2 μl of each antibody/well).After one hour at room temperature (RT) in the dark the cells weretransferred to 96 Deepwell plates. The cells were washed once with 1 mLPBS containing 2% FBS and after re-suspension in 200 μl the samples wereanalyzed on a FACS Calibur flow cytometer. Lymphocytes were gated in theFSC/SSC dot plot according to size and granularity and further analyzedfor expression of CD19, CD3 and activation markers (CD86, CD69). Datawere calculated from dot blots as percentage of cells positively stainedfor activation markers within the CD19+ or CD3+ population using BDCellQest Software.

For assessing the inhibitory property of compounds, compounds were firstdissolved and diluted in DMSO followed by a 1:50 dilution in medium.Splenocytes from Balb/c mice were isolated, re-suspended and transferredto 96 well plates as described above (200 μl/well). The dilutedcompounds or solvent were added to the plates (25 μl) and incubated at37° C. for 1 hour. Then the cultures were stimulated with 25 μl anti-IgMmAb/well (final concentration 30 μg/mL) for 24 hours at 37° C. andstained with anti-mouse CD86-FITC and anti-mouse CD19-PerCP (2 μl ofeach antibody/well). CD86 expression on CD19 positive B cells wasquantified by flow cytometry as described above.

3 Determination of Antibody Production to Sheep Red Blood Cells (SRBC).

In brief, OFA rats were injected i.v. with sheep erythrocytes on dO andtreated orally on four consecutive days (d0 to d3) with the compoundsunder investigation. Spleen cell suspensions were prepared on d4 andlymphocytes were plated onto soft agar in presence of indicator cells(SRBC) and complement. Lysis of the indicator cells due to secretion ofSRBC-specific antibody (predominantly of the IgM subclass) and presenceof complement yielded plaques. The number of plaques per plate werecounted and expressed as number of plaques per spleen.

Immunization:

Groups of five female OFA rats were immunized on day 0 with 2×10⁸/mlSRBC (obtained from Laboratory Animal Services LAS, Novartis Pharma AG)in a volume of 0.5 ml per rat by i.v. injection.

Compound Treatment:

Animals were treated with compound suspended in 0.5% CMC, 0.5% Tween80in for 4 consecutive days (days 0, 1, 2 and 3) starting on the day ofimmunization. Compound was administered orally twice daily with 12 hoursintervals between doses in an application volume of 5 ml/kg body weight.

Preparation of Spleen Cell Suspensions

On day 4, animals were euthanized with CO₂. Spleens were removed,weighed, and deposited in plastic tubes containing 10 ml of cold (4° C.)Hank's balanced salt solution (HBSS; Gibco, pH 7.3, containing 1 mgPhenolred/100 ml) for each rat spleen. Spleens were homogenized with aglass potter, left on ice for 5 minutes and 1 ml supernatant wastransferred into a new tube. Cells were washed once in 4 ml HBSS thensupernatants were discarded and pellets re-suspended in 1 ml of HBSS.Lymphocyte numbers per spleen were determined by automated cell counterand spleen cell suspensions were adjusted to a cell concentration of30×10⁶/ml.

Plaque Forming Assay:

Soft agar petri dishes were prepared with 0.7% agarose (SERVA) in HBSS.

In addition, one ml of 0.7% agarose was prepared in plastic tubes andkept at 48° C. in a water bath. Some 50 μl of a 30×10⁶/ml spleen cellsuspension and 50 μl of SRBC at 40×10⁸/ml were added, mixed rapidly(Vortex) and poured onto the prepared agarose dishes. Petri dishes wereslightly tilted to achieve even distribution of cell mixture on agaroselayer.

The dishes were left at room temperature for 15 minutes and were thenincubated at 37° C. for 60 minutes. Then, 1.4 ml guinea pig complement(Harlan; 10%) was added and the incubation continued for another 60minutes at 37° C. SRBC-specific antibodies released by the plated-out Bcells bound to the antigen (SRBC) in their vicinity. Theseantigen-antibody complexes activated complement and led to the lysis ofthe SRBC leaving a bright spot (plaque) within the red erythrocytelayer. Plaques were counted with a microscope.

The following formula for determination of inhibition of plaqueformation was used:

% Inhibition=C*100/V−100

with: V=mean number of plaques/spleen for vehicle group; C=mean numberof plaques/spleen for compound treated group

REFERENCES

-   N. K. Jerne & A. A. Nordin (1963) Plaque formation in agar by single    antibody-producing cells. Science 140:405.-   N. K. Jerne, A. A. Nordin & C. Henry (1963) The agar plaque    technique for recognizing antibody-producing cells. In: “Cell Bound    Antibodies”, B. Amos & H. Koprowski, Eds., Wistar Inst. Press,    Philadelphia pp. 109-125.

Biological Data

Enzymatic Assay

Example PI3K alpha (uM) PI3K delta (uM) 1 2.0378 0.015 2 3.391 0.009 32.386 0.015 4 1.764 0.033 5 0.749 0.020 6 0.987 0.044 7 1.973 0.013 82.494 0.027 9 2.906 0.009 10 0.668 0.009 11 1.199 0.011 12 0.952 0.01213 1.802 0.013 14 1.832 0.013 15 1.631 0.014 16 1.684 0.016 17 7.6780.017 18 0.871 0.033 19 3.056 0.033 20 1.839 0.048 21 0.320 0.008 220.580 0.008 23 0.129 0.010 24 0.374 0.009 25 0.820 0.026 26 0.368 0.02127 3.410 0.040 28 1.214 0.004 29 2.585 0.011 30 2.831 0.040 31 3.0240.021 32 2.036 0.023 33 1.967 0.018 34 1.648 0.014 35 4.232 0.049 364.103 0.025 37 7.021 0.031 38 3.306 0.016 39 0.434 0.009 40 0.260 0.00641 0.515 0.014 42 0.863 0.013 43 0.728 0.016 44 1.189 0.016 45 0.8600.018 46 0.803 0.027 47 0.656 0.025 48 0.518 0.029 49 0.388 0.034 510.912 0.044 52 1.024 0.046 53 0.504 0.006 54 0.384 0.005 55 0.661 0.00556 0.860 0.013 57 0.590 0.025 58 3.060 0.030 59 9.100 0.028 60 3.3330.045 61 0.589 0.012 62 0.489 0.023 63 0.791 0.051 64 2.331 0.032 650.738 0.023 66 1.280 0.014 67 0.262 0.023 68 0.043 0.007 69 0.056 0.00370 0.121 0.006 71 0.057 0.003 72 0.093 0.004 73 0.054 0.004 74 0.1130.004 75 0.118 0.004 76 0.106 0.007 77 1.290 0.044 78 0.384 0.012 790.781 0.017 80 0.430 0.016 81 0.651 0.02 82 0.066 0.003 83 0.432 0.01784 0.058 0.009 85 0.569 0.021 86 1.330 0.020 87 0.452 0.012 88 1.3360.034 89 1.189 0.029 90 1.991 0.038 91 0.924 0.011 92 2.545 0.009 930.872 0.024 94 1.714 0.021 95 0.757 0.053

Cellular Assays

Cell PI3Kδ/IC50 mCD86/IC50 CD86 Example [umol l−1] [nmol l−1] 1 0.15394.9 5 0.455 125 20 0.2538 120 28 0.268 71.6 65 0.191 570 67 0.047 48.368 0.053 14.9 71 0.035 24.7 81 0.246 86.5 82 0.116 46.2

SRBC Assay

Plaques/spleen Example 1 10 mg/kg bid 12608 ± 4986 Vehicle (0.5% CMC0.5% Tween80) 168363 ± 49142

1-20. (canceled) 21: A method of modulating the activity of PI3Kδisoform in a subject, the method comprising the step of administering toa subject a therapeutically effective amount of a salt form of1-{(S)-3-[6-(6-Methoxy-5-trifluoromethyl-pyridin-3-yl)-5,6,7,8-tetrahydro-pyrido[4,3-d]pyrimidin-4-ylamino]-pyrrolidin-1-yl}-propan-1-one,wherein the salt form comprises an anion selected from phosphate,chloride or hippurate. 22: The method of claim 21, wherein saidmodulating the activity of PI3Kδ isoform in the subject treats Sjögren'ssyndrome in the subject. 23: The method of claim 23, wherein the anionis phosphate. 24: The method of claim 23, wherein the salt is inanhydrous form. 25: The method of claim 24, wherein the salt ischaracterized by an X-Ray powder diffraction pattern comprising thefollowing peaks given at degrees 2-Theta: 11.6, 16.6, 20.7 and 23.3. 26:The method of claim 24, wherein the salt is characterized by an X-Raypowder diffraction pattern comprising the following peaks given atdegrees 2-Theta: 9.8, 11.6, 16.6, 19.5, 20.7 and 23.3. 27: The method ofclaim 22, wherein the anion is chloride. 28: The method of claim 27,wherein the salt is in anhydrous form. 29: The method of claim 28,wherein the salt is characterized by an X-Ray powder diffraction patterncomprising the following peaks given at degrees 2-Theta: 5.6, 11.3 and23.1. 30: The method of claim 28, wherein the salt is characterized byan X-Ray powder diffraction pattern comprising the following peaks givenat degrees 2-Theta: 5.6, 11.3, 14.7, 19.4 and 23.1. 31: The method ofclaim 22, wherein the anion is hippurate. 32: The method of claim 31,wherein the salt is in anhydrous form. 33: The method of claim 32,wherein the salt is characterized by an X-Ray powder diffraction patterncomprising the following peaks given at degrees 2-Theta: 5.2, 7.5, 23.2and 24.2. 34: The method of claim 32, wherein the salt is characterizedby an X-Ray powder diffraction pattern comprising the following peaksgiven at degrees 2-Theta: 5.2, 7.5, 10.3, 10.9, 23.2 and 24.2.